Ion 540 Kit – Chef - Thermo Fisher Scientific · PDF fileNote: Ion 540™ Kit – Chef (Cat. No. A30011), for use when performing two sequencing runs per initialization, only supports

For Research Use Only. Not for use in diagnostic procedures.

Ion 540™ Kit – ChefUSER GUIDE

Instructions for automated template preparation, chip loading,and sequencing

for use with:Ion Chef™ SystemIon S5™ SystemIon S5™ XL System

Catalog Numbers A27759, A30011Publication Number MAN0010851

Revision D.0

The information in this guide is subject to change without notice.DISCLAIMER: TO THE EXTENT ALLOWED BY LAW, LIFE TECHNOLOGIES AND/OR ITS AFFILIATE(S) WILL NOT BE LIABLE FOR SPECIAL, INCIDENTAL,INDIRECT, PUNITIVE, MULTIPLE, OR CONSEQUENTIAL DAMAGES IN CONNECTION WITH OR ARISING FROM THIS DOCUMENT, INCLUDING YOURUSE OF IT.

Revision history: Pub. No. MAN0010851

Revision Date DescriptionD.0 23 January 2017 • “Perform an instrument reset run with an initialized, unused Sequencing

Reagents cartridge“ topic added to Supplemental procedures.

• Designation of Chip-loading centrifuge positions changed from "A" and "B"to "1" and "2"

• ISP quality control sections updated and moved to new user guides:

– Ion Sphere™ Assay on the Qubit™ 2.0 Fluorometer User Guide (Pub.No. MAN0016387)

– Ion Sphere™ Assay on the Qubit™ 3.0 Fluorometer User Guide (Pub.No. MAN0016388)

• Web links updated

C.0 31 July 2016 • Updated to support new Ion S5™ instrument user interface in Torrent Suite™

Software v5.2

• Graphics enhanced and workflows reorganized for ease of use.

B.0 13 November 2015 Update to user guide for 2 sequencing runs per initialization that accompany newSKU (A30011)

A.0 26 August 2015 New user guide, which includes instructions for using the Ion 540™ Kit – Chef toset up and operate Ion Chef™ Instrument for use with the Ion S5™ and Ion S5™ XLSystems

Important Licensing Information: This product may be covered by one or more Limited Use Label Licenses. By use of this product, you accept theterms and conditions of all applicable Limited Use Label Licenses.Corporate entity: Life Technologies Corporation | Carlsbad, CA 92008 USA | Toll Free in USA 1 800 955 6288

Trademarks: All trademarks are the property of Thermo Fisher Scientific and its subsidiaries unless otherwise specified. Microsoft, Windows, andExcel are trademarks of Microsoft Corporation. Macintosh is a trademark of Apple, Inc. Guava and easyCyte are trademarks of Guava Technologies,Inc. Cy is a registered trademark of GE Healthcare UK Limited. Agencourt and Ampure are trademarks of Beckman Coulter, Inc. Bioanalyzer is atrademark of Agilient Technologies, Inc. Kimwipes is a trademark of Kimberly-Clark Corporation.

©2017 Thermo Fisher Scientific Inc. All rights reserved.

Contents

About this guide . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7

Purpose of this guide . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7

Prerequisites . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7

■ CHAPTER 1 Product information . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8

Product description . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8Library compatibility . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8Software compatibility . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8

Ion 540™ Kit – Chef . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9Kit summary . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9Kit contents and storage . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9

Compatible Ion Chip™ kits . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10

Ion S5™ Controls . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10

Required materials not supplied . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11

About the Ion Chef™ System . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12Ion Chef™ Instrument components . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12Interior hardware and consumables . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13About the Ion Chef™ System touchscreen interface . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14

Precautions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14Avoid nucleic acid contamination . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14Avoid chip damage . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14Guidelines for using Ion Chef™ reagents and consumables . . . . . . . . . . . . . . . . . . . . . . 15

Workflow . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 16

■ CHAPTER 2 Before you begin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 17

Create a Planned Run . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 17About Planned Runs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 17Create a Planned Run . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 17Planned Run wizard key fields . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 20

Dilute the libraries . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 22

Prepare the Ion Chef™ System for use . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 23

Ion 540™ Kit – Chef User Guide 3

■ CHAPTER 3 Run the Ion Chef™ System .. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 24

Materials required . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 24

Prepare the libraries and consumables . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 25

Load the Ion Chef™ System . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 26Load the pipette tip racks and PCR plate . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 27Load the Reagents and Solutions cartridges . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 29Load the Recovery Tubes and Enrichment Cartridge v2 . . . . . . . . . . . . . . . . . . . . . . . . . 30Load the Chip-loading centrifuge . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 32Confirm that consumables are correctly installed . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 34Single chip workflow . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 34Ion Chef™ pre-run checklist . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 36

Start the Ion Chef™ run . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 37

Unload the chips for sequencing . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 42

■ CHAPTER 4 Initialize the sequencer . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 45

Ion S5™ System component positions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 45

Before you begin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 46

When a manual cleaning of the sequencer is required . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 46

Initialize the sequencer . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 47

Reagent consumables disposal . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 48CO2 scrubber removal and disposal . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 48Recycle Ion S5™ Wash and Cleaning solution bottles . . . . . . . . . . . . . . . . . . . . . . . . . . . . 49

■ CHAPTER 5 Start the sequencing run . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 50

Chip handling guidelines . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 50

Start the sequencing run . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 51

Maintain the sequencer . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 53Required materials . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 53Clean or decontaminate the sequencer . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 53

■ CHAPTER 6 Clean the Ion Chef™ System . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 54

About the cleaning protocol . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 54

Materials required . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 54

Clean the Ion Chef™ Instrument . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 55Remove and dispose of used consumables . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 56Inspect and clean the Recovery centrifuges and buckets . . . . . . . . . . . . . . . . . . . . . . . . 57Start the cleaning . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 58

Contents

4 Ion 540™ Kit – Chef User Guide

■ APPENDIX A Troubleshooting . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 60

Ion Chef™ System setup and operation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 60

Ion S5™ System setup and operation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 63Ion S5™ Sequencer alarms and events . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 63Initialization—General errors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 64

Ion Sphere™ Assay troubleshooting table . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 65

■ APPENDIX B Supplemental procedures . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 66

Perform a manual cleaning of the sequencer . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 66

Perform an instrument reset run with an initialized, unused SequencingReagents cartridge . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 67

Quality control of Ion 540™ ISPs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 68Quality control using the Ion Sphere™ Assay on the Qubit™ Fluorometer . . . . . . . . . . 68Acceptance criteria for unenriched Ion 540™ ISPs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 68Quality control using the Guava™ easyCyte™ 5 Flow Cytometer . . . . . . . . . . . . . . . . . . . 68Quality control using the Attune™ Acoustic Focusing Cytometer . . . . . . . . . . . . . . . . . . 69

Maintain the Ion Chef™ System . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 69Install a firmware update . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 69Change the instrument name . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 70

■ APPENDIX C Ion Chef™ and Torrent Server network setup . . . . . . . . . 71

Networking overview . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 71About the Ion Chef™ instrument . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 72About the Torrent Server . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 72Networking guidelines and best practices . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 72Network setup workflow . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 72Network setup examples . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 72Collect the required network information . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 72Materials required . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 73General requirements . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 73

Ion Chef™ Networking Configuration Options . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 76Target the Torrent Server IP Address . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 76

Editing the Torrent Server firewall tables to allow the LAN port IP . . . . . . . . . . . . . . . . . . . 80Create an exception . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 80

Enable Planned Run sharing among Torrent Servers . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 82Set up Server Network (admin action) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 83

Transfer a Planned Run . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 84

Undo a Planned Run transfer (administrator) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 86

Undo a Planned Run transfer (user) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 86

Contents


■ APPENDIX D Safety . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 87

Symbols on this instrument . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 88Conformity symbols . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 89

Location of safety labels on this instrument . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 90

Safety information for instruments not manufactured by Thermo Fisher Scientific . . . . . 90

Instrument safety . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 90General . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 90Physical injury . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 90Electrical . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 91Cleaning and decontamination . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 91Ultraviolet (UV) Safety . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 92

Safety and electromagnetic compatibility (EMC) standards . . . . . . . . . . . . . . . . . . . . . . . . . . 92Safety compliance . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 92EMC . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 92Environmental design . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 93

Instrument safety — Ion S5™/Ion S5™ XL System . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 94

Chemical safety . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 94

Biological hazard safety . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 95

■ Documentation and support . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 96

Customer and technical support . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 96

Limited product warranty . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 96

Contents


About this guide

IMPORTANT! Before using this product, read and understand the information in the“Safety” appendix in this document.

Purpose of this guide

This user guide describes how to use the Ion Torrent™ Ion 540™ Kit – Chef (Cat. Nos.A27759, A30011) and the Ion Chef™ System to prepare enriched, template-positive IonSphere™ Particles (ISPs) for up to 200 base-read sequencing of libraries on the Ion S5™

Sequencer or the Ion S5™ XL Sequencer. The kits include all reagents and materialssufficient for performing four dual-sample template-preparation runs on the IonChef™ System for subsequent sequencing on an Ion S5™ System.

WARNING! The protection provided by the equipment may be impaired if theinstrument is operated outside the environment and use specifications, the userprovides inadequate maintenance, or the equipment is used in a manner notspecified by the manufacturer.

Prerequisites

This guide also assumes that you have:• A general understanding of Ion Torrent™ sequencing chemistry and workflow• Knowledge of techniques for handling and preparing DNA libraries


Product information

■ Product description . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8

■ Ion 540™ Kit – Chef . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9

■ Compatible Ion Chip™ kits . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10

■ Ion S5™ Controls . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10

■ Required materials not supplied . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11

■ About the Ion Chef™ System . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12

■ Precautions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14

■ Workflow . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 16

Product description

The Ion 540™ Kit – Chef (Cat. Nos. A27759, A30011) include the reagents andmaterials required to use the Ion Chef™ Instrument to prepare template-positive IonSphere™ Particles and load Ion 540™ Chips. The kit also includes reagents andmaterials to sequence the loaded chips on the Ion S5™ or Ion S5™ XL Sequencer.

Note: Ion 540™ Kit – Chef (Cat. No. A30011), for use when performing twosequencing runs per initialization, only supports 200 base-read sequencing.

The Ion 540™ Kit – Chef can be used with up to 200-base-read libraries of any typeprepared using any available Ion library kit.

The Ion 540™ Kit – Chef is compatible with Torrent Suite™ Software v5.2 and later. Besure to update your Torrent Server to the latest available version of Torrent Suite™

Software before using this kit.

1

Librarycompatibility

Softwarecompatibility


Ion 540™ Kit – Chef

IMPORTANT! Do not substitute components from any other Ion sequencing kits. Wehave verified this protocol using these specific materials. Substitution may adverselyaffect system performance.

IMPORTANT! Store all consumables and cartridges under the recommendedconditions and in an upright position. Do NOT store the Ion S5™ Sequencing Reagents(Cat. No. A27768) on dry ice or in a closed environment where dry ice is present.

Each Ion 540™ Kit – Chef contains all materials that are required to prepare andsequence 8 Ion 540™ Chips.

• Catalog No. A27759 supports 8 initializations• Catalog No. A30011 supports 4 initializations (2 × 200 base-read sequencing runs

per initialization)

On arrival, inspect all consumables and contact Technical Support if any of theproducts have been damaged during shipping.

Component Part No.Quantity per kit

A30011 A27759

Ion S5™ Chef Supplies A27755 4 boxes 4 boxes

Ion S5™ Chef Solutions A27754 1 box 1 box

Ion 540™ Chef Reagents A27758 1 box 1 box

Ion S5™ Sequencing Solutions A27767 1 box 2 boxes

Ion S5™ Sequencing Reagents A27768 1 box 2 boxes

Contents Amount / box Storage

Ion S5™ Chef Supplies (Part No. A27755)

Chip Adapter 2 15°C to 30°C

Enrichment Cartridge v2 1

Tip Cartridge 1

PCR Plate and Frame Seal v2 1 each

Recovery Station Disposable Lid v2 2

Recovery Tube v2 12

Ion S5™ Chef Solutions (Part No. A27754)

Ion S5™ Chef Solutions 4 cartridges 15°C to 30°C

Kit summary

Kit contents andstorage

Chapter 1 Product informationIon 540™ Kit – Chef 1


Contents Amount / box Storage

Ion 540™ Chef Reagents (Part No. A27758)

Ion 540™ Chef Reagents 4 cartridges –30°C to –10°C

Ion S5™ Sequencing Solutions (Part No. A27767)

Ion S5™ Wash Solution 4 × 1.5 L 15°C to 30°C

Ion S5™ Cleaning Solution 250 mL

Ion S5™ Sequencing Reagents (Part No. A27768)

Ion S5™ Sequencing Reagents

(Cartridges ship at 2°C to 8°C. Store asindicated, do not store on dry ice.)

4 cartridges –30°C to –10°C

IMPORTANT! Do not store the Ion S5™ Sequencing Reagents (Part No. A27768) ondry ice or in a closed environment containing dry ice.

Compatible Ion Chip™ kits

Description Catalog No. Quantity Storage

Ion 540™ Chip Kit (2 × 4-pack) A27766 8 chips 15°C to 30°C

Ion S5™ Controls

Human CEPH Genomic DNA Control is sufficient for 3 library preparations (1 µgeach). The Ion S5™ Controls Kit (Cat. No. A27760) is ordered separately.

Contents Amount Storage

Human CEPH Genomic DNA Control (red cap) 30 µL –30°C to –10°C

Human CEPH Control 200 Library (yellow cap) 12 µL

Chapter 1 Product informationCompatible Ion Chip™ kits1


Required materials not supplied

Unless otherwise indicated, all materials are available through thermofisher.com.MLS: Fisher Scientific (fisherscientific.com) or other major laboratory supplier.

Item Source

Non-interruptible Power Supply (UPS) [1] MLS

Thermal cycler with a heated lid MLS

Microcentrifuge[2] MLS

P2, P10, P20, P200, P1000 pipettor set and filtered tips MLS

0.2-mL PCR tubes, flat cap (Axygen™ MAXYMum Recovery™ PCRTube

Fisher Scientific14-222-283

or MLS

Microcentrifuge tubes, 1.5-mL or 1.7-mL MLS

Vortex mixer with a rubber platform MLS

Gloves, powder-free nitrile MLS

Ice buckets and ice —

Nuclease-free water, molecular biology grade MLS

Isopropyl alcohol, 70% solution MLS

Wipes, disposable lint-free MLS

[1] For laboratories that experience frequent power outages or line voltage fluctuations, we recommend that you use a non-interruptible power supply that is compatible with 2500 W output or higher.

[2] Must fit standard 0.2- and 1.5‑mL microcentrifuge tubes and generate 15,500 × g. To convert the RPMs of your centrifuge to RCF in units of gravity, see tools.thermofisher.com/content/sfs/brochures/TR0040-Centrifuge-speed.pdf.

Chapter 1 Product informationRequired materials not supplied 1


http://www.thermofisher.com

http://fisherscientific.com

https://tools.thermofisher.com/content/sfs/brochures/TR0040-Centrifuge-speed.pdf

https://tools.thermofisher.com/content/sfs/brochures/TR0040-Centrifuge-speed.pdf

About the Ion Chef™ System

The Ion Chef™ System provides automated, high-throughput template preparationand chip loading for use with the Ion S5™ or Ion S5™ XL Sequencer. The systemincludes a complete set of cartridge-based consumables and reagents that enable auser to load two Ion 540™ Chips in approximately 13 hours with less than 15 minutesof hands-on time. The Ion Chef™ System features network integration with the TorrentSuite™ Software to enable sample and reagent traceability throughout the chippreparation workflow.

The following figure illustrates the major external and internal features of theIon Chef™ Instrument.

4

1 2 3

5 6 7 8

1 Door – Provides access to the interior ofthe instrument. The door is locked in theclosed position during operation.

2 Micropipettor – A mechanical positive-displacement pipettor that performs allfluid transfers during sample and chippreparation.

3 Robotic arm – Performs fluid transferduring chip loading. The arm alsocontains an optical sensor that reads thebarcodes of instrument reagents andconsumables.

4 Touchscreen – Provides access to allinstrument functions for operation,maintenance, and troubleshooting.

5 Power button – Power switch for theIon Chef™ Instrument, where the statesare on (illuminated) and off.

6 Power port – A 100–240 VAC port thatprovides power to the Ion Chef™

Instrument.7 Ethernet port – An RJ45 port that provides

Ethernet (100 Mbit) communication withthe Ion Chef™ Instrument.

8 USB port – Provides USB communicationwith the Ion Chef™ Instrument. Used toupdate the instrument firmware and totransfer data during service ormaintenance.

Ion Chef™

Instrumentcomponents

Chapter 1 Product informationAbout the Ion Chef™ System1


The following figure illustrates the interior of the Ion Chef™ Instrument and describesthe stations involved in the preparation of chips for sequencing on the Ion S5™ or IonS5™ XL System.

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1 Waste pipette tip rack – The position ofthe rack containing waste (used) pipettetips.

2 Automated heated cover – Transfers theplate cover to the PCR reaction platewithin the sample block. During thermalcycling, the heated cover appliescompression to seal the reaction plateand heats the cover to preventcondensation.

3 New pipette tips – The position of the rackcontaining unused pipette tips.

4 Thermal cycler sample block – Performsthermal cycling of the sequencingreactions on a 96-well PCR reaction plate.

5 Reagents station – The position on theinstrument deck of the diluted libraries,NaOH, and the Ion 540™ Chef Reagentscartridge.

6 Solutions station – The position on theinstrument deck of the Ion S5™ ChefSolutions cartridge, which is maintainedat room temperature.

7 Recovery centrifuges – Twin stations thatperform centrifugation of the ISPs duringthe recovery phase of templatepreparation.

8 Enrichment station – The position of therack containing consumables for theenrichment of the template-positive ISPs.

9 Chip-loading centrifuge – Performscentrifugation of sequencing chips thathave been mounted to chip-loadingadapters and loaded with template-positive ISPs.

Interior hardwareand consumables

Chapter 1 Product informationAbout the Ion Chef™ System 1


The Ion Chef™ System features a simple interface for loading chips, cleaning theinstrument, and performing system maintenance and configuration tasks.

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3 4 5

1 Set up Run button – Set up the Ion Chef™

template preparation and chip-loadingroutine. Choose Step by Step to have theinstrument lead you stepwise throughinstallation of reagents and consumables,or choose Quick Start to proceed if youhave already installed the consumables.

2 Open Door button.3 Notifications button – View notifications

about instrument status during andbetween runs.

4 Quick Start button – Proceed directly tothe Quick Start instrument setup mode.User verifies the loading of a new pipettetip cartridge and an empty pipette tip rackto hold waste tips generated during therun, before proceeding to Deck Scan.

5 Settings button – Advance to a screenproviding the following options:

• Notifications: view notifications aboutinstrument status during and betweenruns

• Instrument Settings: view currentsettings and network configuration,set instrument name, adjust time zone

• Service tools: access screens forservice-related maintenance andinstrument diagnostics

• Torrent Server: add and manageTorrent Server connections

• Clean Ion Chef: proceed directly to theinstrument cleaning routine

• Check for updates: check availabilityof system software updates

Precautions

IMPORTANT! A primary source of contamination is spurious DNA fragments fromprevious sample processing steps. Do not introduce amplified DNA into the librarypreparation laboratory or work area.

IMPORTANT! To avoid possible damage to the chip due to electrostatic discharge,ground yourself before picking up a chip or placing a chip on a surface such as a labbench. For example, touch the metal trim on the chip compartment before inserting orremoving a chip from the chip clamp.

About the IonChef™ Systemtouchscreeninterface

Avoid nucleic acidcontamination

Avoid chip damage

Chapter 1 Product informationPrecautions1


To ensure the proper function of the Ion Chef™ System, handle the associatedconsumables and reagents according to the following guidelines:

• Store all Ion Chef™ System consumables and cartridges under the recommendedconditions and in an upright position.

• Inspect all Ion Chef™ System consumables and cartridges for damage on arrivaland again before use.

• Hold sequencing chips by gently gripping them by their edges.• When the Ion Chef™ System is not in use, remove all consumables and reagents

from the deck and close the instrument door.• Except for the New Pipette Tip Cartridge, do not reuse any of the Ion Chef™

System consumables or reagents. After each run, the empty Pipette Tip Cartridgeis transferred to the waste tip station.

IMPORTANT! All Ion Chef™ Instrument components are single-use only.

• Use only Ion Torrent™ kits and supplies with the Ion Chef™ Instrument. The useof third-party reagents and supplies can adversely affect the performance of theIon Chef™ Instrument and chips prepared.

• Always load two of the same model of sequencing chips into the Ion Chef™

System. The instrument cannot load different models of sequencing chips duringthe same run.

• Remove and sequence chips within 1 hour after the Ion Chef™ System finishesloading them. If you cannot sequence a loaded chip immediately, store it in a chipstorage container at 4°C until you are ready to run it (up to 6–8 hours maximum).

Note: If you choose to store a loaded chip, remove the chip from 4°C storage (butkeep it in the chip storage container) at least 20 minutes before running it,allowing the chip to warm to room temperature.

Guidelines forusing Ion Chef™

reagents andconsumables

Chapter 1 Product informationPrecautions 1


Workflow

The following workflow illustrates how to prepare and load samples using the Ion540™ Kit – Chef, and Ion Chef™ System for sequencing on the Ion S5™ or Ion S5™ XLSequencer.

Create a Planned Run within the Torrent Suite™ Softwareand set the run parameters.

q

Dilute the library samples, and thaw the reagents.

q

Load the Ion Chef™ Instrument with sequencing chips,consumables, reagents, and libraries.

q

Start the Ion Chef™ Instrument

q

Unload the Ion Chef™ Instrument.

q

Load the prepared chip into the Ion S5™ or Ion S5™ XL Sequencer andbegin the sequencing run.

Chapter 1 Product informationWorkflow1


Before you begin

■ Create a Planned Run . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 17

■ Dilute the libraries . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 22

■ Prepare the Ion Chef™ System for use . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 23

Create a Planned Run

Planned Runs contain all the settings used in a sequencing run, including number offlows, kit types, barcodes, sample information, and reference files (if any). They areused to track samples, chips, and reagents throughout the sequencing workflow, fromtemplate preparation on the Ion Chef™ Instrument through sequencing on the Ion S5™

or Ion S5™ XL Sequencer and subsequent data analysis. Each of the two chipsprepared in an Ion Chef™ run requires its own Planned Run.

1. Open the Torrent Browser on a computer connected to your Ion Chef™ System.

2. Select the Plan tab, click Templates, select the application that you want to run(such as AmpliSeq™ Exome), then click either:

• Plan New Run to plan a new run using the generic template for the selectedapplication.

2

About PlannedRuns

Create a PlannedRun


• Plan Run in the dropdown menu under the Settings tab to the right ofthe existing template you select from the template list.

3. In the Planned Run wizard, review the IonReporter and Application tabs andmake selections appropriate to your run. In the Kits tab, make the followingselections:

a. Select Ion S5™ System from the Instrument dropdown list.

b. Select Ion 540™ Chip from the Chip Type dropdown list.

c. Select Ion Chef for Template Kit, and select Ion 540 Kit-Chef from theTemplate Kit dropdown list.

d. Select Ion S5 Sequencing Kit from the Sequencing Kit dropdown list.

Chapter 2 Before you beginCreate a Planned Run2


e. Enter the appropriate number of flows according to the following table:

Sequencing type Number of Flows

200-base-read 500

150-base-read 400

100-base-read 260

f. Select or edit the optional information fields appropriately for your run.

g. Click Next.

4. Review the Plugins and Projects tabs and make selections appropriate to yourrun.

5. In the Plan tab, enter or make the following selections:a. Enter a Run Plan Name and select Reference and BED files appropriate to

your run.

b. Enter the number of chips you are planning runs for. To specify two samplenames, enter "2" in the Number of chips field and then click the check markbutton to the right of this field.

c. Enter a sample name for each plan in the appropriate Sample (required)fields.

d. Enter or scan the barcodes of the Ion Chef™ Library Sample Tubes into theappropriate Sample Tube Label fields.

Note: If you selected a Barcode Set in the Kits tab, fields appear in the Plan tabwhere you enter the number of barcodes used, and the corresponding SampleTube Label. Then enter a sample name for each barcode used in the Samplestable. Return to step 2 and repeat the procedure from the beginning to create a

Chapter 2 Before you beginCreate a Planned Run 2


Planned Run for a second chip. You can copy the first Planned Run in thePlanned Run page and edit appropriately.

Note: For a complete description of the fields of the Torrent Browser application,see the Torrent Suite™ Software Help Guide, available at thermofisher.com/ion-s5-docs.html.

6. When you have completed your selections, click Plan Run at the bottom right ofthe Plan tab screen to save the run. The run is listed on the Planned Runs pageunder the name that you specified and is automatically used by the Ion Chef™

System when the associated sample is loaded.

Field name Description

Ion Reporter If Ion Reporter™ Software is installed and enabled, and you want toanalyze the run data using the software, select the account andworkflow.

Application Select the sequencing application you are performing.

Instrument Select the sequencing system you are using

Chip Type Select the chip type you are using.

Library Kit Type Select the kit used to prepare the library.

Template Kit Select Ion 540 Kit-Chef

Sequencing Kit Select Ion S5 Sequencing Kit

Barcode Set(optional)

If you are using barcodes with:

• DNA libraries: Select the IonXpress barcode set, whichincludes all barcodes in the Ion Xpress™ Barcode Adapters1‑96 Kits.

• RNA libraries prepared using the Ion Total RNA-Seq Kit v2:Select the IonXpressRNA barcode set, which contains all16 barcodes in the Ion Xpress™ RNA BC01-16 Kit(Cat. No. 4475485).

If you are not using barcodes with:

• DNA libraries: Leave the Barcode field blank.

• RNA libraries prepared using the Ion Total RNA-Seq Kit v2:Select RNA_Barcode_None from the dropdown list. This willensure that the proper trimming is performed on theresulting sequence when the RNA library does not have abarcode.

Flows Enter the appropriate number of flows for the read length (forexample, 500 flows for 200‑base‑read sequencing).

Plugins Select the appropriate plugins for your application.

Projects Select or add a project within which to group your run data. Youcan include runs in multiple projects, and remove runs from aproject at any time.

Planned Runwizard key fields

Chapter 2 Before you beginCreate a Planned Run2


http://www.thermofisher.com/ion-s5-docs.html

http://www.thermofisher.com/ion-s5-docs.html

Field name Description

Reference Library Select a reference library uploaded to the Torrent Server, if any.

BED files Select the Target Regions or HotSpot Regions BED file on the IonTorrent Server, if any.

Planned Run Name Enter a name for the run.

Sample Name(Required)

Is Ion Reporter™ Uploader enabled?

• Yes – Enter each sample name and select the appropriatevalues for workflow, relation, relation role, and set ID.

• No – Scan the barcode of the library tube into the "SampleName" and "Sample Tube Label" fields for the specificsample.

Sample Tube Label(Required)

Scan or enter the barcodes located on the Ion Chef™ LibrarySample Tubes that you intend to use to load samples into the IonChef™ Instrument.

IMPORTANT! You must scan or enter the barcode of eachIon Chef™ Library Sample Tube used on the Ion Chef™ Instrument.

MonitoringThresholds

Set thresholds for Bead Loading, Usable Sequence, and KeySignal. In the Torrent Browser Monitor4Runs in Progress tab, analert is displayed if the values for a run fall below the selectedthresholds.

Chapter 2 Before you beginCreate a Planned Run 2


Dilute the libraries

IMPORTANT! Before proceeding, dilute the two Ion libraries to the optimal inputconcentration. The quality of your sequencing data relies greatly upon achieving thecorrect concentration of starting library.

Dilute the two (2) stock libraries with nuclease-free water according to the followingtable. Prepare a fresh dilution of each library before use with the Ion Chef™ System,and use the library dilutions within 48 hours.

Library Recommended concentration [1]

Ion Total RNA-Seq 50−100 pM

Ion TargetSeq™ Exome 50 pM

Ion AmpliSeq™ Exome 50 pM

Ion AmpliSeq™ Exome RDY 50–100 pM

Ion AmpliSeq™ Transcriptome Human GeneExpression 50–100 pM

Ion AmpliSeq™ Comprehensive Cancer Panel 50 pM

Oncomine™ cfDNA Assays 50 pM

Human CEPH Control 200 Library[2] Dilute 1 µL into 24 µL nuclease-freewater

[1] Recommendations are based on qPCR quantification. If libraries are quantified with an Agilent™ 2100 Bioanalyzer™ instrument, a higher calculated concentration may need to be used for equivalent input.

[2] Obtained from the Ion S5™ Controls Kit (Cat. No. A27760).

Chapter 2 Before you beginDilute the libraries2


Prepare the Ion Chef™ System for use

Before you use the Ion Chef™ Instrument:

• Ensure that the Ion Chef™ Instrument has been cleaned following the previousrun. If not, clean the instrument before loading it with consumables.

Note: For more information on the cleaning procedure, see Chapter 6, “Clean theIon Chef™ System“.

• Inspect the empty compartments of the Reagents and Solutions stations forcondensation. Condensate can collect in these compartments, depending ontemperature and humidity conditions. Before loading consumables into theinstrument, wipe the compartments dry with a laboratory wipe or absorbentcloth, if needed.

• Thaw the Ion 540™ Chef Reagents cartridge at room temperature for 45 minutesbefore use.

• Ensure that the Ion Chef™ Instrument has a connection to your Torrent Server. Onthe Ion Chef™ home touchscreen, touch Settings, then Torrent Server to view theconnection status of your instrument.

Note: If the instrument is not connected, see Appendix C, “Ion Chef™ andTorrent Server network setup“ for instructions on how to configure a direct orindirect network connection of the Ion Chef™ Instrument to a Torrent Server.

Chapter 2 Before you beginPrepare the Ion Chef™ System for use 2


Run the Ion Chef™ System

■ Materials required . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 24

■ Prepare the libraries and consumables . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 25

■ Load the Ion Chef™ System . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 26

■ Start the Ion Chef™ run . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 37

■ Unload the chips for sequencing . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 42

This chapter describes:• Setting up the Ion Chef™ Instrument for use by diluting the two Ion libraries, and

loading the instrument with all of the required reagents and consumables• Starting an Ion Chef™ run• Unloading the Ion Chef™ Instrument

Materials required

• Ion 540™ Kit – Chef (Cat. Nos. A27759, A30011)• Ion 540™ Chip (2)• Molecular-biology grade nuclease-free water• P200 pipettor and filtered tips

3


Prepare the libraries and consumables

1. Unbox the Ion 540™ Chef Reagents cartridge 45 minutes prior to use and allow itto warm to room temperature.

IMPORTANT! The Reagents cartridge must sit at room temperature for45 minutes before use.

2. Pipet 25 µL of each diluted library (see “Dilute the libraries“ on page 22) to thebottom of the appropriate Ion Chef™ Library Sample Tube (flagged tubes).

Note: If running the Human CEPH Control 200 Library from the Ion S5™

Controls (Cat. No. A27760), prepare the control library for use by diluting 1 µL ofstock library into 24 µL nuclease-free water.

3. Cap and store the two diluted DNA libraries on ice until you are ready to loadthem onto the Ion Chef™ Instrument.

4. Remove all cartridges and consumables from their wrappings and boxes, thenplace them on the bench next to the Ion Chef™ Instrument.Prepare the following:

• Chip Adapter (2)• Enrichment Cartridge v2• Tip Cartridge v2• PCR Plate and Frame Seal v2• Recovery Station Disposable Lid v2 (2)• Recovery Tube v2 (12)• Ion 540™ Chef Reagents cartridge (from Step 1)• Ion S5™ Chef Solutions cartridge

IMPORTANT! Prior to use, gently tap the Reagents and Solutions cartridges onthe bench to force the reagents to the bottoms of the tubes.

Note: When stored under normal conditions, a precipitate can form in sometubes of the Ion 540™ Chef Reagents cartridge. If present, load the cartridge asdirected – the precipitate will dissolve when the reagents are mixed duringinstrument operation.

Chapter 3 Run the Ion Chef™ SystemPrepare the libraries and consumables 3


Load the Ion Chef™ System

IMPORTANT!· Rated centrifuge speeds are only intended for operation with the provided buckets

and approved consumable chips, tubes, and sample preparation reagents.· The Chip-loading centrifuge is rated to operate at the listed rotational frequencies

with the chip buckets, chips, and adapters. The centrifuge must be load-balanced.Proper care must be taken to load the buckets properly. If excessive vibrations arise,check that items are installed properly and rotors are load-balanced.

· Use only the materials supplied in the Ion 540™ Kit – Chef to run the centrifuges atthe rated speeds. Do not remove or change the rotors. Inspect the buckets beforeeach use to assure normal operation.

· Confirm that the instrument is powered ON and has been cleaned following thelast use.

· Ensure all components are clean and dry before loading them onto the Ion Chef™

Instrument.· Ensure the Reagents and Solutions station compartments are free of condensate

before loading components.

Follow the procedure below to load the Ion Chef™ Instrument. A completely loadedinstrument is shown in the following figure:

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1 Empty tip rack (move from new TipCartridge position)

2 Frame Seal v23 New Tip Cartridge4 PCR plate5 Ion 540™ Chef Reagents cartridge

6 Ion S5™ Chef Solutions cartridge7 Recovery Tubes and Recovery Station

Disposable Lid v28 Enrichment Cartridge v29 Chip Adapter/Chip assemblies

Chapter 3 Run the Ion Chef™ SystemLoad the Ion Chef™ System3


1. Touch (Open Door) in the instrument touchscreen to open the instrumentdoor, then wait for the latch to open.

2. Lift the instrument door to the top of the travel until the latch mechanismengages.

1

1 Hold here, then lift

3. Load an empty pipette tip rack to the Used (Waste) Pipette Tip Position, thenchange gloves.

11

1 Used Pipette Tip Position

IMPORTANT!· Confirm that the pipette tip rack in the Used (Waste) Pipette Tip Position does

not contain any tips. The instrument will abort the run if tips are present in theused position.

· To prevent contamination, change gloves immediately after moving the emptypipette tip rack to the Used (Waste) Pipette Tip Position.

Note: A small amount of dried residue may be present in the tub of the emptypipette tip rack after a run. This will not affect the next run in the Used PipetteTip Position.

4. Unwrap a new Tip Cartridge and remove the cover to expose the pipette tips,then load it in the New Pipette Tip Position.

Note: Two Ion Chef™ Piercing Tips are pre-loaded into tip positions G7 and H7on the Tip Cartridge.

Load the pipettetip racks and PCRplate

Chapter 3 Run the Ion Chef™ SystemLoad the Ion Chef™ System 3


5. Slide the catch forward to allow the locking bracket to pivot upward. Load theTip Cartridge into the New Pipette Tip Position, pull the bracket downward, thenpush the catch backward to lock the bracket and cartridge in place.

21

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1 New Pipette Tip Position2 Bracket3 Catch4 New Tip Cartridge

6. Load a new PCR plate into the thermal cycler sample block, then slide a newFrame Seal v2 under the automated heated cover.

IMPORTANT! When the Frame Seal v2 is positioned correctly, its tabs projectupward and contact the heated cover.

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1 Thermal cycler sample block2 Well A13 Cover4 Keyed corner



IMPORTANT! Thaw the Reagents cartridge at room temperature for 45 minutesbefore use.

1. Gently tap the Ion 540™ Chef Reagents cartridge on the bench to force thereagents to the bottoms of the tubes.

2. Load the cartridge into the Reagents station so that it snaps into place and is levelon the deck.

IMPORTANT! Do not force the Ion Chef™ cartridges into place. Each cartridgefits only one location on the deck and in one orientation. If a cartridge does notfit, confirm that you are loading the correct cartridge in the correct orientation.

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1 Reagents station (4°C)2 Ion 540™ Chef Reagents cartridge

3. Uncap, then load the two Library Sample Tubes, each containing 25 µL of dilutedlibrary, into Positions A and B on the Reagents cartridge.

A

B00012647

000122161

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C 3

D 4

1 Position A (Library)2 Position B (Library)3 Position C (NaOH)4 Position D (Empty tube)

Load the Reagentsand Solutionscartridges



IMPORTANT!· Make sure to orient the sample tubes so that the barcodes are visible and

oriented to the right.· Make sure to remove the caps to the Library Sample Tubes before proceeding.· Because 200- and 400-base-read libraries require different run lengths, do not

load a 200-base-read library and 400-base-read library in a single Ion Chef™

run. Both libraries loaded in a run must have a similar read length.

4. Uncap both the tube of NaOH in Position C and the empty tube in Position D onthe Reagents cartridge.

IMPORTANT! When the Reagents cartridge is loaded:

· Press down on the Library Sample Tubes to ensure that they are firmly seatedin the cartridge.

· Confirm that all tubes are uncapped, including the tube at Position D.

5. Gently tap the Ion S5™ Chef Solutions cartridge on the bench to force the reagentsto the bottoms of the tubes.

6. Load the Solutions cartridge into the Solutions station until it snaps into placeand is level on the deck.

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1. Load six Recovery Tubes (v2) into each Recovery centrifuge.

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Load the RecoveryTubes andEnrichmentCartridge v2



Before sealing each centrifuge, confirm that:• The centrifuge is load-balanced with all required consumables.

IMPORTANT! The centrifuge must be load-balanced.

• The buckets are securely seated in the centrifuge rotors.• The buckets are oriented correctly in the centrifuge so that they pivot

outwards.

2. Place a Recovery Station Disposable Lid v2 over each centrifuge by lining up thetab with the depression on the deck, then snap into place. Ensure that the lidssnap completely into place by applying firm downward pressure along the lidperimeter.

3. Close the hinged cover of the Recovery centrifuges. Confirm that the port of eachdisposable lid is positioned toward the rear of the instrument.

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1 Recovery Tubes installed2 Recovery Station Disposable Lids installed3 Recovery centrifuge cover closed4 Port

IMPORTANT!· Do not obstruct or place any object on top of the Recovery centrifuge cover.· Use only the supplied materials, including buckets and disposables, to run the

centrifuges at the rated speeds. Do not remove or change the rotors. Inspectthe buckets to assure normal operation before each use.



4. Load the Enrichment Cartridge v2, then press down on the cartridge to ensurethat it is level with the instrument deck.

IMPORTANT! Confirm that the Enrichment Cartridge v2 is loaded so that thelettering on the cartridge is right-side-up.

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1. Load each prepared sequencing chipinto a centrifuge bucket, then attacha Chip Adapter to the assembly.

a. Place the chip in the chip-loading bucket with the keyedcorners of the chip and bucketaligned, then align the wells ofthe Chip Adapter to the wells ofthe chip, orienting the adapteronto the chip so that the chipbarcode is visible.

b. Place the adapter onto the chip,then insert the stationary tabs atthe reservoir end of the adapterinto the slots of the bucket.

c. Gently squeeze the flexible tabs at the other end of the adapter into thebucket slots until the adapter locks into place.

d. Confirm that the tabs at all four corners of the adapter are fitted into theslots in the centrifuge bucket. Loading can fail if the adapter is not attachedsecurely.

Note: If desired, you can label the tops of chips to distinguish them. Do notobstruct or overwrite the chip barcode with your label.

Load the Chip-loading centrifuge

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1 Chip Adapter2 Ion chip3 Bucket4 Ports (align with chip)5 Flexible tabs6 Keyed corner (align with bucket)7 Slots



2. Load the adapter/chip/bucket assemblies into the Chip-loading centrifuge.

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IMPORTANT! When the Chip-loading centrifuge is loaded, confirm that eachChip Adapter is firmly attached to a bucket, and that the buckets are securelyseated in the centrifuge rotors.

540CCBD02226

540 CCBD02226

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1 Chip barcode2 Chip position 13 Position 1 marker hole4 Chip position 25 Position 2 marker hole

Note: Position 1 of the Chip-loading centrifuge is the position 90° clockwisefrom the single hole in the rotor bucket cover at rest. The chip loaded in Position1 will be loaded with ISPs prepared from the DNA library in the Library SampleTube loaded in Position A of the Reagents cartridge. The chip loaded in Position2 of the centrifuge will be loaded with ISPs prepared from the DNA library in theLibrary Sample Tube loaded in Position B of the Reagents cartridge.



3. Ensure the centrifuge is load-balanced, and the chip buckets are securely seatedand oriented correctly in the centrifuge so that they pivot 90° outwards whentouched. Then close the lid of the Chip-loading centrifuge.

IMPORTANT! Do not obstruct or place any object on top of the lid.

Before continuing:• Confirm that each cartridge is at the correct location and in the correct

orientation.• Press down on all cartridges to confirm that they are firmly pressed into place.• Confirm that all tubes in the Ion 540™ Chef Reagents cartridge, including the tube

of NaOH in Position C, are uncapped and firmly pressed into place.• Confirm that the centrifuge lids are installed correctly so that the port is oriented

toward the rear of the instrument.• Confirm that the tube and chip buckets are seated securely in the rotor arms of

the Chip-loading and Recovery centrifuges, and that the consumables theycontain are correctly installed.

CAUTION! To ensure correct and safe instrument operation, you must confirmthat all consumables are installed correctly to the deck before you start a run.The Ion Chef™ Instrument does not verify all aspects of the consumable setupprior to beginning each run.

You can set up an Ion Chef™ run to load asingle chip instead of two, using theIon Chef™ S5 Series Chip Balance loadedopposite to the sequencing chip in theChip-loading centrifuge. The Ion Chef™

S5 Series Chip Balance is provided in theIon S5™ Installation Kit.

Load the Ion Chef™ Instrument as youwould normally load the system. Forsingle chip loading, perform thefollowing steps:

1. Add the single diluted DNA library to an Ion Chef™ Library Sample Tube andload the tube into Position A of the Reagents cartridge.

2. Load an empty Ion Chef™ Library Sample Tube into Position B of the Reagentscartridge. Uncap both tubes.

3. Load the chip in Position 1 and the Ion Chef™ S5 Series Chip Balance in Position 2of the Chip-loading centrifuge.

IMPORTANT! Do not use Ion Chef™ 314, 316/318, or P-Series versions of the chipbalance with the Ion 540™ Chip. Each chip balance is weight-matched to the chip(and corresponding chip adapter) specified on the chip balance label.

Confirm thatconsumables arecorrectly installed

Single chipworkflow

Ion Chef™ S5 Series

Chip Balance

Ion Chef™ S5 Series Chip Balance



Note: Position A of the Chip-loading centrifuge is the position 90° clockwisefrom the single hole in the rotor bucket cover at rest.

540DABD02226

Ion Chef™S5 Series

Chip Balance 1 3

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1 Position 1 (chip)2 Position 2 marker holes3 Position 2 (Chip Balance)4 Position 1 marker holes

4. Resume the normal workflow in “Load the Chip-loading centrifuge“ at step 3.The Ion Chef™ Instrument will detect the presence of the single chip during DeckScan before the run starts.



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Ion Che f ™ S5 Series Chip Balance

1) A new Ion Chef™ Tip Cartridge v2 is loaded in the New Pipette Tip position.2) Bracket is pulled downward to lock the tip cartridge in place.

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1) A new PCR Plate is loaded in the thermal cycler sample block.2) A Frame Seal v2 is in place and oriented under the heated cover. 1) The tip rack from previous run is transferred

from the New Pipette Tip position to the Used Pipette Tip position.

1) Recovery Tubes (v2) are securely seated in the Recovery centrifuge buckets.2) Recovery centrifuges are load-balanced.3) Recovery Station Disposable Lids (v2) are positioned so that the port is correctly oriented: port should not be located in the position facing the front of the instrument.

1) Enrichment Cartridge v2 is pressed into place at the Enrichment station so that the cartridge is firmly seated and level with the deck.2) Lettering on the cartridge is right-side up and positioned to the right of the enrichment tubes.1) Adapter/chip assembly is correctly

seated in the Chip-loading centrifuge buckets, with the adaptor clips inserted into the bucket slots.2) Centrifuge buckets containing the adapter/chip assemblies are securely seated in the centrifuge rotor, and freely pivot 90º outwards.3) Chip-loading centrifuge is load-balanced.

1) Solutions cartridge is pressed into place at the Solutions station so that the cartridge is firmly seated and level with the deck.

1) Reagents cartridge contents are thawed (leave at room temperature for 45 minutes).2) Reagents cartridge tubes are uncapped and pressed into place at the Reagents station so that the tubes are firmly seated and level with the deck.3) Library Sample tubes are firmly seated and in the correct orientation with barcodes facing the PCR Plate and caps removed.

Ion Chef™ pre-run checklist



Start the Ion Chef™ run

1. Confirm that you have loaded the instrument with all kits and consumables.

2. On the Ion Chef™ Instrument home touchscreen, touch Set up run.

3. Touch Step by Step to have the instrument lead you through the instrumentsetup, or touch Quick Start to skip the instrument setup screens.

Chapter 3 Run the Ion Chef™ SystemStart the Ion Chef™ run 3


4. Follow the on-screen instructions. When prompted, close the instrument door byfirst lifting it slightly to disengage the locking mechanism, then push down onthe door until the locks engage. After the door closes, the instrument visionsystem activates.

IMPORTANT! Do not close the door by pulling it straight down from the openposition. You must lift the door slightly before you can close it. Ensure that bothsides of the door are locked after closing it.

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1 Lift door first2 Lower3 Press down to lock

5. When prompted, touch Start check to start Deck Scan. Wait while the instrumentscans the barcodes of all consumables and reagents to confirm their presence andcompatibility.During Deck Scan, the touchscreen displays warnings if the Ion Chef™

Instrument detects missing or incompatible consumables. You must address allwarnings before the run can start. After you address each condition, touch Yes tocontinue.

IMPORTANT! The Deck Scan function is not a substitute for manual inspectionof the reagents and consumables on the Ion Chef™ Instrument before starting arun. To ensure proper and safe instrument operation, confirm that allconsumables are installed correctly before you continue.

6. When Deck Scan is complete, touch Next to display the Data Destination screen.

Chapter 3 Run the Ion Chef™ SystemStart the Ion Chef™ run3


7. Confirm that the instrument displays the correct kit type, chip type, chipbarcodes, and Planned Run. If the correct Planned Runs do not display, touch thedropdown menu to select the Planned Run for each chip, then touch Next.

IMPORTANT! If the kit and chip type are not correct, confirm that you are usingthe correct kit and chip. If you are using the correct kit and chip, and an incorrectkit or chip type is displayed on the screen, contact Technical Support.

8. On the Run Options screen, touch the appropriate option to complete the run,then enter the desired time of run completion, if needed.

The Ion Chef™ Instrument provides two options for obtaining quality control(QC) samples that can be used to evaluate templating efficiency. Depending onyour selection, the QC samples are made available either during (before chiploading) or after the run. In either case, you can obtain unenriched samples fromthe corresponding Library Sample Tubes at Positions A and B on the Reagentscartridge, or enriched samples from Positions A and E on the EnrichmentCartridge v2.

By selecting You can obtain the QC samples

Time immediately after the run ends, at the time you specify(12 hours, 45 minutes after run start).

Pause when the instrument pauses operation before the chiploading step (about 11 hours into the run).

Note: The DNA library in the Library Sample Tube loaded in Position A of theReagents cartridge is templated onto ISPs that can be sampled in Position E ofthe Enrichment Cartridge v2 after a run. The DNA library in the Library Sample



Tube loaded in Position B of the Reagents cartridge is templated onto ISPs thatcan be sampled in Position A of the Enrichment Cartridge v2.

Note: Select Pause if you are uncertain of library quality and want to evaluatetemplating efficiency before chips are loaded. If you do not pause the run, youcan collect QC samples after the run. Save the samples until sequence analysis iscomplete to have them available for troubleshooting.

9. On the Run Options screen, touch Start run to begin the run.

Note: If you need to stop the run for any reason, touch Cancel, then touch Yes toconfirm the cancellation.

If the Ion Chef™ Instrument encounters a problem during the run, it aborts therun and displays the error on the instrument touchscreen. If a run fails:

a. Remove the consumables from the deck, then clean the instrument. Ifpossible, retain the consumables for troubleshooting.

b. Reset, then reattempt the run. If the run fails again, contact TechnicalSupport to troubleshoot the problem.

10. Initialize the Ion S5™ or Ion S5™ XL Sequencer at least 40 minutes before theIon Chef™ System finishes chip loading.Initialization of the instrument can be performed up to 24 hours before starting asequencing run. If the intent is to perform two sequencing runs per initialization,the first run must be completed and the second run started within the 24-hourperiod. By initializing the sequencer before completion of chip loading, youensure that the chips can be sequenced as soon as possible after loading iscomplete.

11. If you chose to pause the run to analyze the templating efficiency, remove thesamples for testing when prompted to do so by the Ion Chef™ Instrument(approximately 11 hours after the start of the run).

a. When prompted to remove the QC sample, open the instrument door.

IMPORTANT! If you unintentionally close the instrument door before youobtain the QC samples, you must wait until the end of the run before youcan collect them. You cannot pause the run or open the door after it has beenclosed.

Chapter 3 Run the Ion Chef™ SystemStart the Ion Chef™ run3


b. Transfer the QC samples (entire volume) from Positions A and B of the Ion540™ Chef Reagents cartridge on the instrument deck to two new labeledmicrocentrifuge tubes.

IMPORTANT! Do not remove the Library Sample Tubes from the Ion 540™

Chef Reagents cartridge.

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00012647

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1 Position A (QC Sample)2 Position B (QC Sample)

c. Analyze the QC samples.

Note: For detailed protocols, see the· Ion Sphere™ Assay on the Qubit™ 2.0 Fluorometer User Guide (Pub. No.

MAN0016387) or· Ion Sphere™ Assay on the Qubit™ 3.0 Fluorometer User Guide (Pub. No.

MAN0016388)

available at thermofisher.com/order/catalog/product/4468656.

d. If you are performing quality assessment of enriched samples, transfer QCsamples from positions A and E of the Enrichment Cartridge v2 to two newlabeled microcentrifuge tubes. See “Quality control ofIon 540™ ISPs“ onpage 68.

e. Close the instrument door, then touch Continue to complete the run.

12. When the run is complete, unload the Ion Chef™ Instrument and sequence thechips immediately. You can collect QC samples from the Reagents and/orEnrichment cartridges if you have not done so already.

Note: If you cannot sequence a loaded chip immediately, place the chip into aseparate chip storage container and store at 4°C until you are ready to sequenceit (up to 6–8 hours maximum).



http://www.thermofisher.com/order/catalog/product/4468656

Unload the chips for sequencing

1. Open the instrument door:a. In the instrument touchscreen, touch (Open Door) then wait for the latch

to open.

b. Lift the instrument door to the top of the travel until the latch mechanismengages.

1


2. Open the lid of the Chip-loading centrifuge, then unload both chip/bucketassemblies from the Ion Chef™ Instrument.

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To unload each chip:a. Remove the chip/bucket assembly from the Chip-loading centrifuge.

b. Apply pressure to both ends of the Chip Adapter, then remove and discardthe Chip Adapter.

Chapter 3 Run the Ion Chef™ SystemUnload the chips for sequencing3


c. Grasp the chip by its edges, carefully lift the chip out of the bucket, then setit aside on a clean, static-free surface. Return the bucket to the Chip-loadingcentrifuge.

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1 Chip Adapter2 Ion chip3 Bucket4 Press and remove adapter5 Remove from bucket carefully

3. Close the instrument door by first lifting it slightly to disengage the lockingmechanism, then push down on the door until the locks engage.

IMPORTANT! Do not close the door by pulling it straight down from the openposition. You must lift the door slightly before you can close it. Ensure that bothsides of the door are locked after closing it.

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1 Lift door first2 Lower3 Press down to lock

Chapter 3 Run the Ion Chef™ SystemUnload the chips for sequencing 3


4. Load one or both chips into Ion S5™ Sequencers and promptly start thesequencing runs.If you cannot sequence a loaded chip immediately or plan to sequence two chipsper initialization, place the chip into a separate chip storage container and storeat 4°C until you are ready to sequence it (up to 6–8 hours maximum).

IMPORTANT! If you chose to store a loaded chip, remove the chip from thecontainer and place it on a clean surface in the dark to warm to roomtemperature at least 20 minutes before running it.

Chapter 3 Run the Ion Chef™ SystemUnload the chips for sequencing3


Initialize the sequencer

■ Ion S5™ System component positions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 45

■ Before you begin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 46

■ When a manual cleaning of the sequencer is required . . . . . . . . . . . . . . . . . . . . . 46

■ Initialize the sequencer . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 47

■ Reagent consumables disposal . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 48

Initialization takes ~40 minutes.

Ion S5™ System component positions

7

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1 Touchscreen2 Power button3 Ion S5™ Sequencing Reagents cartridge4 Chip clamp

5 Ion S5™ Wash Solution bottle. Wastereservoir located behind the WashSolution bottle (shown on the right).

6 Ion S5™ Cleaning Solution bottle7 Waste reservoir

Note: The system uses RFID technology to verify that the proper reagents are loadedin positions 3, 5, and 6. Reagents that exceed their expiration date or usage countgenerate an error message prompting the user to replace the reagent beforeperforming the run.

Note: RFID regulatory information can be found on the main screen underOptions4Regulatory info.

4


Before you begin

The Ion S5™ and Ion S5™ XL Sequencers are equipped to verify the compatibility ofeach chip and consumable loaded during initialization and sequencing, and that thesecomponents do not exceed their expiration date. To avoid exceptions duringinitialization, inspect this information for each consumable before installing onto theinstrument.

• Unbox the Ion S5™ Sequencing Reagents cartridge 45 minutes before use, thenallow it to equilibrate to room temperature.Do not remove the Ion S5™ Sequencing Reagents cartridge from its foil packaginguntil immediately before loading, so that you can return the unused cartridge tostorage if your sequencing run is delayed.

• Unbox the Ion S5™ Wash Solution bottle. Invert the bottle 5 times, then swirl at anangle to mix thoroughly.

• Remove the red cap from the Ion S5™ Wash Solution and Ion S5™ CleaningSolution bottles immediately before installing on the instrument.

When a manual cleaning of the sequencer is required

The Ion S5™ Sequencer and Ion S5™ XL Sequencer require that a cleaning beperformed before initialization. This is normally performed automatically at thecompletion of the previous sequencing run. When two sequencing runs areperformed on a single initialization, the post-run cleaning is performed after thesecond sequencing run. However, if the "Enable post-run clean" checkbox (see “Startthe sequencing run“) is unchecked to allow a second run, and a second run is notperformed, the instrument will not allow the subsequent initialization to proceeduntil a manual cleaning has been performed. See “Perform a manual cleaning of thesequencer“ on page 66 for more information on how to perform a manual cleaning.

If an Ion S5™ Sequencer or an Ion S5™ XL Sequencer is initialized and a sequencingrun is not started within 24 hours, or a run is not started or completed due to a powerfailure or an abort, do not perform a manual cleaning. An instrument reset run isrequired before reinitialization. See “Perform an instrument reset run with aninitialized, unused Sequencing Reagents cartridge“ on page 67 for moreinformation.

Chapter 4 Initialize the sequencerBefore you begin4


Initialize the sequencer

1. In the instrument touchscreen main menu, touch Initialize. The door, chip, andReagent cartridge clamps unlock.

2. When prompted, remove the Ion S5™ Wash Solution bottle to access the wastereservoir, then remove and empty the waste reservoir.

3. Reinstall the empty waste reservoir.

4. Replace the expended Ion S5™ Sequencing Reagents cartridge with a newcartridge equilibrated to room temperature.

5. Ensure the new Ion S5™ Wash Solution bottle is thoroughly mixed. Then removethe red cap and install.

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Chapter 4 Initialize the sequencerInitialize the sequencer 4


6. Ensure that the used sequencing chip from the previous run is properly seated inthe chip clamp and the chip clamp is pushed in all the way.

7. If necessary, install a new Ion S5™ Cleaning Solution bottle.

Note: The Ion S5™ Cleaning Solution bottle contains sufficient reagent tocomplete 4 cleanings.

8. Close the door, then touch Next.The instrument confirms that the consumables and chip are properly installedand that the Ion S5™ Cleaning Solution bottle contains sufficient reagent toperform the post-run clean. Follow all on-screen recommendations to ensureproper installation of required consumables.

IMPORTANT! If the allowed number of post-run cleans has been met, theinstrument prompts the user to replace the Ion S5™ Cleaning Solution bottle.

Note: See “Bottle leak check fails“ on page 64 if a Checking Reagent: Failedwarning appears.

9. When initialization is complete (~30−40 minutes), touch Home.

The instrument is now ready for a sequencing run.

Reagent consumables disposal

IMPORTANT! Follow all applicable local, state/provincial, and/or national regulationswhen recycling or disposing of Ion S5™ reagent consumables.

To properly dispose of the CO2 scrubber, you must first remove it from the Ion S5™

Sequencing Reagents cartridge.

1. Invert the Ion S5™ Sequencing Reagents cartridgeover an appropriate receptacle to drain allresidual liquid.

2. Wearing gloves, insert the Ion S5™ Cartridge Toolfirmly into the CO2 scrubber until the flangestops on the top of the cartridge.

CO2 scrubberremoval anddisposal

Ion S5™ Cartridge Tool

Chapter 4 Initialize the sequencerReagent consumables disposal4


3. Pull straight up on the tool while holding the nucleotide reagent cartridge down.

4. Remove the scrubber from the cartridge tool, then dispose of the scrubberaccording to applicable hazardous waste regulations.

The Ion S5™ Wash solution and Ion S5™ Cleaning solution bottles are made ofrecyclable plastic.

1. Open the expended bottle by unscrewing the cap.

2. Remove the cap, sipper, and filter, then pour any residual liquid into anappropriate receptacle.

3. Rinse the empty bottle with water. Pour out the rinse water into the same liquidwaste receptacle.

4. Recycle or dispose of the clean bottle, cap, sipper, and filter according toapplicable regulations.

Recycle Ion S5™

Wash andCleaning solutionbottles

Chapter 4 Initialize the sequencerReagent consumables disposal 4


Start the sequencing run

■ Chip handling guidelines . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 50

■ Start the sequencing run . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 51

■ Maintain the sequencer . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 53

Chip handling guidelines

To place a chip in the chip clamp:

1 2 3 4

1 Slide the chip clamp out.2 Remove the chip currently in the clamp.3 Place the appropriate loaded chip in the

chip clamp with the chip notch in thebottom ‑front corner.

Note: Do not force the chip into theclamp. If the chip does not fit easily in theclamp, confirm that the notch is orientedas shown in the drawing.

4 Slide the metal tab in fully to engage theclamp, then close the instrument door.

5


Start the sequencing run

We recommend that you sequence loaded chips on the Ion S5™ or Ion S5™ XLSequencer as soon as possible after chip loading and instrument initialization arecomplete. However, successful sequencing runs can be started up to 24 hours afterinstrument initialization.

Note: Do not press the power button during a run. Interrupting power to theinstrument during a run may result in sequencing run failure and loss of sample.

1. After completion of initialization, press Run in the instrument touchscreen. Thedoor and chip clamp unlock.

2. Remove the used sequencing chip, then secure a chip loaded with template-positive Ion Sphere™ Particles in the chip clamp.

3. Push the chip clamp all the way in to engage, close the instrument door, thenpress Next.

4. Confirm that the correct Planned Run has auto-populated, then press Review.

5. (Optional) If this is to be the first of two sequencing runs on this initialization,deselect the Enable post-run clean checkbox, then press Review.

Note:· Failure to deselect the checkbox results in a cleaning performed automatically

after the first run. A second run is not available.· When starting the second sequencing run on a single initialization, ensure the

Enable post-run clean checkbox is selected so that the post-run cleaning isperformed automatically.

Chapter 5 Start the sequencing runStart the sequencing run 5


6. Confirm the remaining pre-populated settings are correct, or make changes usingthe buttons and dropdown lists if necessary.

7. Confirm that the instrument door is closed, then press Start run to begin thesequencing run.

IMPORTANT! During a run, do not open the instrument door, and avoidtouching the instrument. Touching the instrument during the sequencing runmay reduce the quality of the measurements.

When the sequencing run is complete, the instrument automatically performs thecleaning procedure unless the Enable post-run clean checkbox was deselected.After cleaning, the touchscreen returns to the main menu. Use the TorrentBrowser to review the results.

Chapter 5 Start the sequencing runStart the sequencing run5


Maintain the sequencer

• Lint-free wipes• 70% isopropanol• (optional)– 10% bleach solution

In the event of a spill or leak on or inside the instrument, perform the following steps.

Note: Dispose of all waste in appropriate liquid or solid waste containers.

1. Remove the Ion S5™ Wash Solution bottle, then remove and empty the wastereservoir.

2. Remove the Ion S5™ Sequencing Reagents cartridge.

3. Inspect the waste and nucleotide reagent bays for liquid.

4. Using absorbent paper soak up as much liquid as possible, then wash theaffected area with 10% bleach solution.

5. Wipe the affected surfaces with 70% isopropanol, then allow to air-dry.

Requiredmaterials

Clean ordecontaminate thesequencer

Chapter 5 Start the sequencing runMaintain the sequencer 5


Clean the Ion Chef™ System

■ About the cleaning protocol . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 54

■ Materials required . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 54

■ Clean the Ion Chef™ Instrument . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 55

About the cleaning protocol

The Ion Chef™ System includes an automated cleaning function that must beperformed following every run. The cleaning routine is initiated from the Ion Chef™

Instrument touchscreen and is designed to minimize potential contamination. Duringthe routine, the instrument irradiates the deck with ultraviolet light for 1 minute afterall consumables have been removed from the instrument.

IMPORTANT! Although the Ion Chef™ Instrument cleaning routine provides someprotection against contamination, it is not a substitute for good laboratory techniqueor precautions. When preparing DNA libraries for use or when preparing theIon Chef™ Instrument, make certain to observe sterile laboratory procedures at alltimes to ensure minimal contamination.

Materials required

• Gloves, powder-free nitrile• Isopropanol, 70% solution• Wipes, lint-free

6


Clean the Ion Chef™ Instrument

IMPORTANT! Clean the Ion Chef™ Instrument as described in the following pagesafter every run. To prevent contamination, do not operate the instrument unless it hasbeen recently cleaned.

4

5

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2 1

8 7

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Ion Chef™ Instrument stations1 Waste pipette tip position2 Empty Tip Cartridge: move to waste

pipette tip station3 Thermal cycler sample block4 Reagents station

5 Solutions station6 Recovery centrifuges7 Enrichment station8 Chip-loading centrifuge

Chapter 6 Clean the Ion Chef™ SystemClean the Ion Chef™ Instrument 6


IMPORTANT!· Do not discard the empty Tip Cartridge.· Make sure to transfer the QC samples before you remove and discard the Reagents

cartridge.

1. Touch (Open Door) in the instrument touchscreen, then wait for the latch toopen.

2. Lift the instrument door to the top of the travel until the latch mechanismengages.

1


3. Remove, then discard the PCR plate from the thermal cycler sample block.

4. Remove, then discard the box of used pipette tips from the waste tip position.

IMPORTANT! Handle the disposable reservoir in the waste tip position withcare. During the run, liquid waste collects in the reservoir. Dispose of the liquidwaste by tipping the reservoir on one corner and pouring the waste into anappropriate waste container:

IMPORTANT! Do not reuse the waste pipette tip rack. Always move the emptyTip Cartridge from the new tip position to the waste tip position.

5. Move the empty Tip Cartridge to the waste tip position.

Remove anddispose of usedconsumables

Chapter 6 Clean the Ion Chef™ SystemClean the Ion Chef™ Instrument6


6. Remove, then discard the• Ion 540™ Chef Reagents cartridge• Ion S5™ Chef Solutions cartridge• Enrichment Cartridge v2

7. Remove, then discard the consumables from the Recovery centrifuges, includingthe:

• Recovery Station Disposable Lids v2• Recovery Tubes v2

8. Close the Chip-loading centrifuge cover.

1. Inspect the Recovery centrifuges, then clean the components if excess liquid ispresent.

Is liquidpresent? Action

No Go to “Start the cleaning“ on page 58.

Yes Clean the centrifuge bowl and buckets as described below.

IMPORTANT! Clean the Recovery centrifuges occasionally, onlywhen excess liquid is noticeable in the bowl and/or buckets. Youdo not need to clean the centrifuge after every run.

IMPORTANT! Wear powder-free, nitrile gloves when cleaning the Recoverycentrifuge.

2. Remove the buckets from the Recovery centrifuges. Clean the inside and outsideof each bucket using a lint-free wipe, then place the buckets on a clean, drysurface while you clean the centrifuge.

1

2

1 Bucket2 Lint-free wipe

Inspect and cleanthe Recoverycentrifuges andbuckets



3. Use lint-free wipes to remove all fluid from inside the centrifuge bowl.

1

2

1 Inside rim of the centrifuge2 Bottom of the centrifuge bowls

4. Use lint-free wipes treated with 70% isopropanol to clean the:• Inside rim of the centrifuge.• Bottom of the centrifuge bowl.• Outside and inside of the centrifuge buckets.

5. Dry the centrifuge and buckets with lint-free wipes.

6. Install the centrifuge buckets, then close the Recovery centrifuge cover.

1

1 Buckets (cleaned and installed)

1. Close the instrument door by first lifting it up slightly to disengage the lockingmechanism, then pushing down on the door until the locks engage.

IMPORTANT! Before closing the door, ensure the covers of the centrifuges areclosed.

Start the cleaning

Chapter 6 Clean the Ion Chef™ SystemClean the Ion Chef™ Instrument6


2. To start the cleaning, touch Next on the Ion Chef™ Instrument touchscreen thatappears after run completion.

Note: You can also clean the instrument at any time starting from the hometouchscreen. Touch Settings4Clean Ion Chef.

3. Confirm that you have removed all consumables from the Ion Chef™ Instrument,except the empty pipette tip rack in the waste tip position, then touch Next.

4. With the door closed, touch Start. The instrument performs a Deck Scan beforestarting the cleaning routine. The Ion Chef™ Instrument stops ventilation, thenilluminates the ultraviolet (UV) light in the instrument for ~1 minute.

CAUTION! The Ion Chef™ Instrument emits UV light at 254 nm. Wearappropriate eye wear, protective clothing, and gloves when working nearthe instrument. Do not look directly at the UV light while it is illuminatedduring the cleaning routine.



Troubleshooting

■ Ion Chef™ System setup and operation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 60

■ Ion S5™ System setup and operation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 63

■ Ion Sphere™ Assay troubleshooting table . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 65

Ion Chef™ System setup and operation

Observation Possible cause Recommended action

Instrument does not display thehome screen when powered on

Multiple causes are possible. 1. Power off the instrument, wait30 seconds, then power on theinstrument.

2. If the instrument fails again, contactTechnical Support.

Cannot open the instrumentdoor

An obstruction is present on oraround the door mechanism.

Remove the obstruction blocking the door, thenoperate the instrument normally.

A hardware or software errorhas occurred.

Contact Technical Support to report theproblem and for further assistance.

Instrument stops during a run The instrument hasencountered an internal error.

1. Record the error displayed on theinstrument display, then touch OK.

2. Contact Technical Support to report theproblem and for further assistance.

Liquid residue is present in theRecovery centrifuges followinga run

During normal instrumentoperation, a noticeable coatingof liquid can collect on the bowland buckets of the Recoverycentrifuges following repeatedruns.

Remove the residue as instructed in “Inspectand clean the Recovery centrifuges andbuckets“ on page 57.

A



Instrument will not begin a run The instrument hasencountered a Deck Scan error(one or more consumables areabsent or loaded improperly).

1. Confirm that the touchscreen does notdisplay any Deck Scan warnings. If alarmsare present, note the error(s) displayed,replace the missing consumable asdirected, press No when prompted thenpress Next to cancel the run. Afterreturning to the home screen, restart therun.

2. If the error persists, confirm that:

• All buckets are seated correctly inthe rotors of the Recovery and Chip-Loading Centrifuges.

• All cartridges are loaded correctlyand are level on the instrument deck.

• The barcodes of the Ion Chef™

Library Sample Tubes are visible andpositioned correctly.

• All tubes are both present anduncapped on the Ion 540™ ChefReagents (Library Sample Tubes,NaOH tube, and the empty tube).

3. If the error persists after you check theconsumables on the instrument deck, doone of the following:

• If you are confident that the Ion Chef™

Instrument is set up correctly andyou are comfortable disregarding thewarnings, touch YES following DeckScan to proceed with the run.

• If the instrument cannot begin therun, contact Technical Support forfurther assistance.

The instrument hasencountered an internal error.

1. Record the error displayed on theinstrument display, then touch OK.

2. Contact Technical Support to report theproblem and for further assistance.

Appendix A TroubleshootingIon Chef™ System setup and operation A



Instrument displays one ormore alerts during a run

The instrument has detectedone or more problems duringthe run.

After the instrument completes the run,contact Technical Support.

IMPORTANT! The detected problem mightimpact the performance of the sequencing run.

• Network connection to theserver has beeninterrupted.

• User name or password isincorrect.

1. Touch the Instrument status button toview the alert(s).

2. In the Instrument status screen, confirmthat the name of the Torrent Serverconnection is red.

3. Contact your network administrator toconfirm that:

• The Torrent Server can be accessedfrom the network port used by theIon Chef™ Instrument. If not,troubleshoot the network connection.

• The user name and password usedby the Ion Chef™ Instrument arevalid. If not, contact the serveradministrator to renew thecredentials.

4. If the alert persists, contact TechnicalSupport for further assistance.

Appendix A TroubleshootingIon Chef™ System setup and operationA


Ion S5™ System setup and operation


Red "Alarms" and/or "Events" message in MainMenu

• Softwareupdatesavailable

• Connectivityissues

• Instrument notdetectingrequired filesor hardware

Click on the red pop-up to see detailedmessages.

• If a message states "NewerSoftware Available":

IMPORTANT! After updates areinstalled, the instrument must berestarted.

a. On the home screen, selectSettings4Check for Updates.

b. Select the Released Updatescheckbox, then press Update.

c. When installation is complete,follow the onscreen promptsto restart the instrument.

Note: In some cases, theinstrument restartsautomatically after softwareinstallation.

• If a message states "NoConnectivity to Torrent Server", "NoConnectivity to ftp server", or"Network Manager not connected",disconnect and re-connect theethernet cable, confirm that therouter is operational, and verifythat the network is up and running.

• For any other messages:a. Power off the instrument: On

the home screen, selectSettings4System Tools4ShutDown4Shut Down.

b. Wait 30 seconds, then pressthe button on the front ofinstrument to power on theinstrument.

• If the red "Alarms" and/or "Events"message still appears in the mainmenu, contact Technical Support.

Ion S5™

Sequencer alarmsand events

Appendix A TroubleshootingIon S5™ System setup and operation A



Bottle leak check fails • Bottle seal is not tight.

• Bottle may be damaged /defective.

1. Finger-tighten the bottles.2. If the bottle continues to leak, replace the

bottle.3. If leak check continues to fail, contact

Technical Support.

Initialization—General errors

Appendix A TroubleshootingIon S5™ System setup and operationA


Ion Sphere™ Assay troubleshooting table

The following table contains troubleshooting information for unenriched ISPs testedwith the Ion Sphere™ Assay on the Qubit™ 2.0 or Qubit™ 3.0 Fluorometer.

Qubit™

Fluorometerobservation

Ion S5™ Systemobservation Possible cause Recommended action

<10%Templated

ISPs

• Lower loading

• Lower % enriched

• Lower key signal

• Lower throughput

Too little library input intotemplate preparation

• Increase library input to target 20–25% templated ISPs.or

• Continue with sequencing; expectlower throughput.

>30%Templated

ISPs,

but <70%

Increased number offiltered reads

Too much library input intotemplate preparation

• Decrease library input to target 20–25% templated ISPs.or

• Continue with sequencing; expectlower throughput.

>70%Templated

ISPs

• Increased % primerdimer filtered reads


Adapter dimercontaminating library, morelikely in short amplicon, IonAmpliSeq™ or miRNAlibraries

• Check Bioanalyzer™ traces foradapter dimer peak (Amplicon libraryor Ion AmpliSeq™ library peak around70 bp; miRNA library peak around60bp).

• Re-purify library using Agencourt™

AMPure™ XP Kit clean-up steps asoutlined in the appropriate userguides.

• Low loading

• Low % enriched


• High % filtered reads

Ion Chef™ System underperformance

Troubleshoot with Technical Support or aField Application Scientist.

Appendix A TroubleshootingIon Sphere™ Assay troubleshooting table A


Supplemental procedures

■ Perform a manual cleaning of the sequencer . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 66

■ Perform an instrument reset run with an initialized, unused SequencingReagents cartridge . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 67

■ Quality control of Ion 540™ ISPs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 68

■ Maintain the Ion Chef™ System . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 69

Perform a manual cleaning of the sequencer

A cleaning protocol is normally performed automatically at the completion of eachsequencing run. In the event that a cleaning is necessary, follow the listed steps:

1. On the home screen, select Settings4Clean Instrument.The instrument door unlocks allowing access to the consumables.

2. Remove the Ion S5™ Wash Solution bottle to access the waste reservoir, thenremove and empty the waste reservoir.

1

2

3

3. Reinstall the empty waste reservoir and the used Ion S5™ Wash Solution bottle.

4. Ensure the Ion S5™ Sequencing Reagents cartridge and Ion S5™ Cleaning Solutionbottle are properly installed.

IMPORTANT! Perform the cleaning with a used reagent cartridge and washsolution bottle installed. The cleaning procedure pumps cleaning solution intothe wash solution bottle and reagent cartridge making them unsuitable forsequencing.

5. Place a used sequencing chip in the chip clamp, then push the chip clamp in allthe way to engage.

B


6. Close the instrument door, then press Next.Cleaning takes ~35 minutes to complete. Upon completion the instrument doorautomatically unlocks and the chip and cartridge clamps disengage.

7. Proceed to “Initialize the sequencer“ on page 47.

Perform an instrument reset run with an initialized, unusedSequencing Reagents cartridge

Cleaning is normally performed at completion of a sequencing run automatically. Ifan Ion S5™ Sequencer or an Ion S5™ XL Sequencer is initialized and

• a sequencing run is not started within 24 hours after initialization, or• a sequencing run is not completed due to a power failure or an abort, and <200flows occurred before the stoppage

an instrument reset run is required to ensure proper cleaning before reinitialization.Do NOT perform a manual cleaning with an unused, initialized Ion S5™ SequencingReagents cartridge.

Note:· If a power failure or abort occurs during the second of two runs started after a

single initialization, a manual cleaning (page 66) is sufficient.· If the number of flows that occurred before a power failure or abort is unknown,

perform an instrument reset run.

To perform an instrument reset run, use the following procedure beforereinitialization:

1. In the instrument touchscreen main menu, press Run.The instrument door and chip clamp unlocks.

2. Ensure that a used sequencing chip is in the chip clamp, then push the chipclamp in all the way to engage.

3. Close the instrument door, then press Next.

4. When prompted, select Planned Run (none). Ensure that the Enable post-runclean checkbox is selected, then press Review.

5. In the Select Run screen, press Edit, then in the Detail screen set the number offlows to 200 manually. Ensure that the Post-Run/Clean checkbox is selected, thenpress Close.

6. Press Start run, then press Accept to confirm that Post-Run Clean is enabled, andto start the run.

When the instrument reset run completes, the instrument automatically performs thecleaning procedure. After cleaning, the touchscreen returns to the main menu.

Appendix B Supplemental proceduresPerform an instrument reset run with an initialized, unused Sequencing Re-

agents cartridgeB


Quality control of Ion 540™ ISPs

The Qubit™ 2.0 Fluorometer or the Qubit™ 3.0 Fluorometer can be used to perform aquality assessment of unenriched Ion Sphere™ Particles generated for up to 200 base-read sequencing on the Ion S5™ Sequencer. For detailed protocols, see the

• Ion Sphere™ Assay on the Qubit™ 2.0 Fluorometer User Guide (Pub. No.MAN0016387) or

• Ion Sphere™ Assay on the Qubit™ 3.0 Fluorometer User Guide (Pub. No.MAN0016388)

available at thermofisher.com/order/catalog/product/4468656.

The optimal amount of library corresponds to the library dilution point that givesPercent Templated ISPs between 10–30%.

Samples that fall within the recommended range generally produce the most data;however, samples that fall outside of the recommended range can still meet thethroughput specifications on the Ion chips.

The recommended optimal range is not intended to be a pass/fail criteria. The rangeprovides guidance for the quality of the sample.

Note: If the results are outside the desired Percent Templated ISPs range, thenincrease or decrease the library input appropriately. See the “Ion Sphere™ Assaytroubleshooting table“ on page 65 for more information.

Percent Templated ISPs Description

<10% Sample contains an insufficient number of templated ISPsto achieve optimal loading density on the Ion Chip.

10–30% Optimal amount of library.

>30% Sample will yield multi-templated ISPs (mixed reads).

The Guava™ easyCyte™ 5 Flow Cytometer can be used for quality assessment ofunenriched and enriched Ion Sphere™ Particles generated for up to 200 base-readsequencing on the Ion S5™ Sequencer or Ion S5™ XL Sequencer.

• Unenriched samples – Obtain the QC samples from the corresponding LibrarySample Tubes on the Ion 540™ Chef Reagents cartridge (positions A and B).

• Enriched samples – Obtain sample 1 from position E and sample 2 from positionA on the Enrichment Cartridge v2.

For details, see the Ion PI™ Ion Sphere™ Particles Quality Assessment Using the Guava™

easyCyte™ 5 Flow Cytometer User Bulletin (Pub. No. MAN0007496), available at thermofisher.com.

Quality controlusing the IonSphere™ Assay onthe Qubit™

Fluorometer

Acceptancecriteria forunenriched Ion540™ ISPs

Quality controlusing the Guava™

easyCyte™ 5 FlowCytometer

Appendix B Supplemental proceduresQuality control of Ion 540™ ISPsB


http://www.thermofisher.com/order/catalog/product/4468656


The Applied Biosystems™ Attune™ Acoustic Focusing Cytometer can be used forquality assessment of unenriched and enriched Ion Sphere™ Particles generated for upto 200 base-read sequencing on the Ion S5™ Sequencer or Ion S5™ XL Sequencer.

• Unenriched samples – Obtain the QC samples from the corresponding LibrarySample Tubes on the Ion 540™ Chef Reagents cartridge (positions A and B).

• Enriched samples – Obtain sample 1 from position E and sample 2 from positionA on the Enrichment Cartridge v2.

For details, refer to the Demonstrated Protocol: Ion Sphere™ Particles (ISPs) QualityAssessment using the Applied Biosystems™ Attune™ Acoustic Focusing Cytometer UserBulletin (Pub. No. 4477181), available at thermofisher.com.

Maintain the Ion Chef™ System

The Ion Chef™ System supports the user-performed maintenance procedures found inthis section.

To ensure proper operation of the Ion Chef™ Instrument, we recommend periodicallyconfirming that your instrument is running the most current firmware. Weoccasionally release updates to the instrument firmware, which can include importantchanges to the Ion Chef™ System operation. To ensure that your instrument is runningthe most current firmware, check the firmware version as explained below.

Check the Ion Chef™ Instrument firmware

1. On the Ion Chef™ Instrument touchscreen, touch Settings.

2. On the Settings screen, touch Check for updates.

3. On the Software Update screen, touch Release to search the server for updates tothe Ion Chef™ Instrument firmware.

Note: Users are also notified of the availability of firmware updates on theNotifications screen.

4. Select the available component update, touch Update, then wait for theinstrument to complete the update.

Note: If the Ion Chef™ Instrument is operating on an isolated network andcannot connect to our website, you must transfer the firmware update manually.

Quality controlusing the Attune™

Acoustic FocusingCytometer

Install a firmwareupdate

Appendix B Supplemental proceduresMaintain the Ion Chef™ System B



To perform a manual update, transfer the firmware files to a USB drive and insertthe drive into the USB port at the rear of the instrument. Touch USB on theSoftware Update screen, select from the list of available software components,then touch Update.

When finished, the Ion Chef™ Instrument displays the update status.

5. Power off and on the Ion Chef™ Instrument to complete the update.

IMPORTANT! If you are updating the firmware manually, remove the USB drivebefore powering on the Ion Chef™ Instrument.

The following procedure describes how to change the name used to identify the IonChef™ Instrument both on the network and in the data that it generates.

1. In the Ion Chef™ Instrument home screen, touch Settings.

2. In the Settings screen, touch Instrument settings.

3. In the Instrument settings screen, touch Set Instrument Name. Touch the namefield and enter a new instrument name using the keypad that appears, thentouch Save.

Note: Use only alphanumeric characters. Do not use special characters or spacesin the name.

4. In the next screen, touch Save again. Power off and on the Ion Chef™ Instrumentto effect the name change.

Change theinstrument name

Appendix B Supplemental proceduresMaintain the Ion Chef™ SystemB


Ion Chef™ and Torrent Servernetwork setup

■ Networking overview . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 71

■ Ion Chef™ Networking Configuration Options . . . . . . . . . . . . . . . . . . . . . . . . . . . 76

■ Editing the Torrent Server firewall tables to allow the LAN port IP . . . . . . . . . 80

■ Enable Planned Run sharing among Torrent Servers . . . . . . . . . . . . . . . . . . . . . . 82

■ Transfer a Planned Run . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 84

■ Undo a Planned Run transfer (administrator) . . . . . . . . . . . . . . . . . . . . . . . . . . . . 86

■ Undo a Planned Run transfer (user) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 86

Networking overview

If you are preparing a laboratory for use with new Ion instruments, refer to therespective site preparation and safety guides. If you currently have an Ion Chef™

instrument configured in your laboratory to communicate to more than one TorrentServer, Torrent Suite™ Software v4.4 or later requires the configuration of theIon Chef™ instrument to communicate with one primary Torrent Server. Afterupgrading to Torrent Suite™ Software v4.4 or later, follow the guidelines in thisdocument to configure the instrument to communicate with a primary Torrent Server.From that Torrent Server, the new Planned Run sharing option can be used to targetadditional Torrent Servers if needed.

Networking with the Torrent Server running Torrent Suite™ Assay DevelopmentSoftware and the Ion PGM™ Dx System requires v5.0 or later. The Ion Chef™ will notconnect to the Ion PGM™ Dx in IVD mode.

If you need to connect a Torrent Server to the network for the first time, see theTorrent Server networking requirements in the site prep and safety guide for therespective sequencer.

This guide describes how to:• Set up the Ion Chef™ instrument to talk to the Torrent Server on a local area

network (LAN).• Enable Planned Run sharing.

C


The instrument is factory-configured for IPv4 TCP/IP communication and includes afast Ethernet adapter (1000 Mbps) with a RJ45-type connector for integrating thedevice into a local area network (LAN). If the instrument will be connected to a LAN,an active network port must be available/assigned in place before the scheduledinstallation date.

If you are activating a newly installed LAN port for your Ion Chef™ instrument,please ensure the LAN has been configured to permit HTTP-443, SSH-22, andFTP-20/21 traffic. For additional information, please refer to the NetworkingRequirements section of the Ion Chef™ Site Preparation Guide (Pub. no. MAN0007956).

The Torrent Server is an integral part of the system and includes a quad-port gigabitNIC for direct communication with the network or instruments such as Ion Chef™,PGM™, Ion Proton™, or Ion S5™ instruments. When the Torrent Server is connected tothe network, Torrent Servers that are on the same software versions and subnet cantransfer planned runs from one Torrent Server to another.

We recommend that you consult a network administrator before connecting theIon Chef™ instrument to your laboratory network. Please refer to the networkingrequirements in the Ion Chef™ System Site Preparation Guide.

Collect the required network information.

q

Connect the Ion Chef™ instrument to the configured LAN port.q

Configure the Ion Chef™ instrument with the required information.q

Monitor the Ion Chef™ instrument to test the network connection.

This guide provides a basic description of how to integrate the Torrent Server and theIon Chef™ instruments into several possible network architectures. Because yournetwork may contain advanced features, such as a firewall or network domains, werecommend that you consult a network administrator before connectingIon Chef™instrument to your laboratory network.

Network examples include:• Direct connection• Static IP (indirect connection)• DHCP IP (indirect connection)

Obtain the following information from your network administrator:• Network policy for obtaining IP addresses (DHCP IP or static IP).• If the network requires static IP addresses, obtain the static IP address, subnet

mask, gateway address, and DNS server name for the LAN port.• For the Planned Run sharing and FTP functionality to be successful, the subnet

must be the same on all Torrent Servers and Ion Chef™ instruments on the LAN.

About theIon Chef™

instrument

About the TorrentServer

Networkingguidelines andbest practices

Network setupworkflow

Network setupexamples

Collect therequired networkinformation

Appendix C Ion Chef™ and Torrent Server network setupNetworking overviewC


An ethernet cable with RJ45 connectors (CAT6 Ethernet cable for a 1000Mbit/snetwork connection or a CAT5 for 100Mbit/s connection).

See Ion Chef™ System Site Preparation Guide (Pub. no. MAN0007956) for additionalinformation.

See Ion Chef™ System Site Preparation Guide (Pub. no. MAN0007956).

DHCP IP

Ion Chef

Network

DHCP Server

Torrent Server

If using DHCP for the LAN port, please make sure the port is set to auto-populatewhen the Ion Chef™ instrument is connected and ensure that all the belowrequirements are met.

Materials required

Generalrequirements

Appendix C Ion Chef™ and Torrent Server network setupNetworking overview C


Static IP

Ion Chef

Network

Torrent Server

If the network requires static IP addresses, obtain the static IP address, subnet mask,gateway address, and DNS server name for the LAN port.

For planned run sharing and FTP functionality to be successful, the subnet must bethe same on all Torrent Servers and Ion Chef™ instruments on the LAN.

Direct connection

Ion Chef

Network

Torrent Server

To configure the direct connection between an Ion Chef™ instrument and a TorrentServer, you will need a Category 6 cable of sufficient length.

Appendix C Ion Chef™ and Torrent Server network setupNetworking overviewC


Another direct connection configuration option consists of one Ion Chef™ instrument,one Torrent Server and two Ion S5™ XL Sequencers. However, this configuration is notrecommended due to increased analysis times.

WARNING! The direct configuration shown below is known to result inlengthy analysis times.

Ion Chef

Network

Torrent Server

S5 XL 1 S5 XL 2

Appendix C Ion Chef™ and Torrent Server network setupNetworking overview C


Ion Chef™ Networking Configuration Options

With the Torrent Suite™ software v4.4 release, the Ion Chef™ software will onlyconnect to one Torrent Server. From that server, Planned Run sharing can beconfigured to access additional servers. When the Ion Chef™ is connected to a TorrentServer, an icon on the lower left side of the screen appears.

Connect the Ion Chef™ Instrument to the Torrent Server in one of three ways: Direct,Static IP (non-direct) or DHCP IP (non-direct). See respective instructions below.

To target the Torrent Server IP address from the Ion Chef™user interface, you will firstlook up the Torrent Server IP address and then enter it into the Ion Chef™userinterface as the Torrent Browser target and the Update Target.

1. Log into Torrent Browser as the admin.

2. Click the Gear button and select Configure.

Target the TorrentServer IP Address

Appendix C Ion Chef™ and Torrent Server network setupIon Chef™ Networking Configuration OptionsC


3. Scroll down to the Database Administration section and click the AdminInterface link.

Appendix C Ion Chef™ and Torrent Server network setupIon Chef™ Networking Configuration Options C


4. The Site Administration screen appears. Scroll to the bottom and click ViewNetwork Settings.

Appendix C Ion Chef™ and Torrent Server network setupIon Chef™ Networking Configuration OptionsC


5. Note (copy or write down) the Torrent Server IP address displayed on theNetwork Settings screen.

6. Go back to the Ion Chef™ user interface and tap on the Torrent Browser IPaddress. This opens the Manage Connections window.

Appendix C Ion Chef™ and Torrent Server network setupIon Chef™ Networking Configuration Options C


7. Enter the server IP address under Server and repeat for Update Site. Click Save.

Editing the Torrent Server firewall tables to allow the LAN port IP

In addition, when connecting through a LAN, the Torrent Server firewall settings willneed to be modified for both DHCP and Static LAN connections.

Note: While directly connected, instruments are on the 192.168.20x.x subnet fromwhich all traffic is enabled, so this is not required.

Any exception would mean that editing the local firewall tables to include the LAN IPis required. Also, if the Ion Chef™ instrument is getting a DHCP address from yournetwork, you will need to create an exception for the entire DHCP subnet.

1. On the Torrent Server, edit the /etc/iptables.custom file and add a firewall rule toallow all traffic from Ion Chef™ instrument's IP address as shown below. (This isa blank file initially and is intended for custom firewall configuration.)

IMPORTANT! Only edit the /etc/iptables.custom file and not /etc/iptables.rules,since /etc/iptables.rules is overwritten when software is updated.

2. Enter "sudo^nano^/etc/iptables.custom" and add the three lines below:• *filter• -A INPUT -s xxx.xxx.xxx.xxx -j ACCEPT• COMMIT

Create anexception

Appendix C Ion Chef™ and Torrent Server network setupEditing the Torrent Server firewall tables to allow the LAN port IPC


Note: -A INPUT -s xxx.xxx.xxx.xxx -j ACCEPT adds the rule to theformat. Enter the IP address of the Ion Chef™ instrument in place of"xxx.xxx.xxx.xxx". When finished editing, save changes by pressing CTRL +O and CTRL + X.

3. Follow this command by manually applying the change:sudoîptables-restore^--noflush^<^/etc/iptables.customNote: You can also list the firewall rules with: sudoîptables-L.

If this command returns with no output, it means it succeeded.

4. If the Ion Chef™ instrument is getting a DHCP address from your network,please create the exception for the entire DHCP subnet. The example belowassumes that the DHCP address for the Ion Chef™ instrument is 172.16.48.0 andthe DHCP subnet range is 172.16.48.1 to 172.16.48.254. Add these 3 lines:

• *filter• -A INPUT -s 172.16.48.0/24 -j ACCEPT• COMMIT

5. Reboot the server.

6. If you want to repeat this process and completely drop all rules and start fresh,enter:

• sudoîptables-F• sudoîptables-restore^<^/etc/iptables.rules• sudoîptables-restore--noflush^</êtc/iptables.custom

Removing a rule from either of these files would remove it from the activefirewall rules, because the iptables-restore command does not overwriterules; it appends them to all active rules.

Appendix C Ion Chef™ and Torrent Server network setupEditing the Torrent Server firewall tables to allow the LAN port IP C


Enable Planned Run sharing among Torrent Servers

New in Torrent Suite™ v4.4, Planned Runs created on one Torrent Server can now betransferred to another Torrent Server if the sequencer connected directly to the serveris offline or busy. Possible configurations include:

Ion Chef™ instrument 1 → Torrent Server 1 ↔ Torrent Server 2 → Sequencer 2

Ion Chef™ instrument 1 → Torrent Server 1 ↔ Torrent Server 3 → Sequencer 3

Network

TS 1(origin)

IC 2IC 1 S5 XL 1 S5 XL 2 IC 3

TS 2(destination)

TS 3(destination)

S5 XL 3

Requirements include:• All Torrent Servers must be running the same software version.• All Torrent Servers must have the same genomic reference, barcode set, BED files,

Variant Caller config files, etc. All Torrent Servers must be on the same subnet.

Appendix C Ion Chef™ and Torrent Server network setupEnable Planned Run sharing among Torrent ServersC


1. On the origin server (e.g., TS1) Site administration page, scroll down and selectShared servers. The Select shared server to change window appears.

2. If you are adding your destination server for the first time, click Add sharedserver.

3. Define your destination server.a. Enter the name, address (can be IP address), user name and password for

the destination server.

b. Click Active if you want this server enabled for sharing.

c. (Optional) Add a comment.

d. Click one of the Save options.

4. (Optional) If you want to configure the origin Torrent Server to also be adestination server, you must go to another server and repeat these steps to set theorigin server as a destination server. Once the Torrent Servers are configured, youor a user can now transfer Planned Runs between Torrent Servers.

Set up ServerNetwork (adminaction)

Appendix C Ion Chef™ and Torrent Server network setupEnable Planned Run sharing among Torrent Servers C


Transfer a Planned Run

You can execute a Planned Run on the Ion Chef™ instrument connected to the originserver, and then, when the run is complete, transfer the Planned Run to a destinationserver for sequencing.

Alternatively, you can create the Planned Run on an origin server and then transfer itto a destination server for both the Ion Chef™ run and the sequencing run.

1. Using the Torrent Browser on the origin Torrent Server, go to Plan4Planned RunList.

2. Open the Gear menu of the Planned Run you want to transfer, and selectTransfer. Then select the destination Torrent Server.

3. A confirmation window appears. Check the information, then click Transfer.

Note: You can no longer access this Planned Run on the origin server after it hastransferred.

Appendix C Ion Chef™ and Torrent Server network setupTransfer a Planned RunC


4. A status window displays the results of the transfer:• The green box lists the samples successfully processed and the required

target BED files found on the destination server.• The red box lists any required BED files or plugins that are not present on

the destination server. To successfully perform the run, you will need to editthe transferred Planned Run on the destination server and manually add themissing BED files or plugins.

5. To edit the transferred Planned Run and add missing files:a. Download required files using the References tab of the Torrent browser of

the destination server

b. Go to the Edit Plan wizard of the transferred Planned Run by selecting Editon the gear pull-down menu to the right of the Planned Run.

c. Select the files or plugins as needed, then click Update Plan.

Note: You can also navigate to the Edit Plan wizard by clicking the Edit testplan link in the status page above.

Appendix C Ion Chef™ and Torrent Server network setupTransfer a Planned Run C


Undo a Planned Run transfer (administrator)

1. On the destination server, delete the transferred Planned Run from either thePlanned Run page or the admin page.

2. On the origin server, locate the plan onthe /admin/rundb/plannedexperiment/page, uncheck PlanExecuted and changePlanStatus to Planned.

Undo a Planned Run transfer (user)

If you transferred a Planned Run in error, you can transfer it back to the origin serveror to another server.

1. On the destination Torrent Server, navigate to Plan4Planned Run List andlocate the transferred Planned Run.

2. From the Gear menu of the Planned Run, select Transfer, then select theTorrent Server to which you wish to transfer the run.

Appendix C Ion Chef™ and Torrent Server network setupUndo a Planned Run transfer (administrator)C


Safety

■ Symbols on this instrument . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 88

■ Location of safety labels on this instrument . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 90

■ Safety information for instruments not manufactured by ThermoFisher Scientific . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 90

■ Instrument safety . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 90

■ Safety and electromagnetic compatibility (EMC) standards . . . . . . . . . . . . . . . . 92

■ Instrument safety — Ion S5™/Ion S5™ XL System . . . . . . . . . . . . . . . . . . . . . . . . 94

■ Chemical safety . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 94

■ Biological hazard safety . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 95

WARNING! GENERAL SAFETY. Using this product in a manner not specifiedin the user documentation may result in personal injury or damage to theinstrument or device. Ensure that anyone using this product has receivedinstructions in general safety practices for laboratories and the safetyinformation provided in this document.

· Before using an instrument or device, read and understand the safetyinformation provided in the user documentation provided by themanufacturer of the instrument or device.

· Before handling chemicals, read and understand all applicable Safety DataSheets (SDSs) and use appropriate personal protective equipment (gloves,gowns, eye protection, etc). To obtain SDSs, see the “Documentation andSupport” section in this document.

D


Symbols on this instrument

Symbols may be found on the instrument to warn against potential hazards or conveyimportant safety information. In this document, the hazard symbol is used along withone of the following user attention words:

• CAUTION! – Indicates a potentially hazardous situation that, if not avoided,may result in minor or moderate injury. It may also be used to alert againstunsafe practices.

• WARNING! – Indicates a potentially hazardous situation that, if not avoided,could result in death or serious injury.

• DANGER! – Indicates an imminently hazardous situation that, if not avoided,will result in death or serious injury.

Symbol English Français

Caution, risk of danger

Consult the manual for further safetyinformation.

Attention, risque de danger

Consulter le manuel pour d’autresrenseignements de sécurité.

Moving parts Parties mobiles

Caution, hot surface Attention, surface chaude

Ultraviolet light Rayonnement ultraviolet

On/Off On/Off (marche/arrêt)

Standby En attente

Protective conductor terminal (mainground)

Borne de conducteur de protection(mise à la terre principale)

Terminal that can receive or supplyalternating current or voltage

Borne pouvant recevoir ou envoyerune tension ou un courant de typealternatif

Appendix D SafetySymbols on this instrumentD


Symbol English Français

Do not dispose of this product inunsorted municipal waste

CAUTION! To minimizenegative environmentalimpact from disposal ofelectronic waste, do notdispose of electronic waste inunsorted municipal waste.Follow local municipal wasteordinances for properdisposal provision andcontact customer service forinformation about responsibledisposal options.

Ne pas éliminer ce produit avec lesdéchets usuels non soumis au trisélectif.

CAUTION! Pour minimiserles conséquences négativessur l’environnement à la suitede l’élimination de déchetsélectroniques, ne pas élimin-er ce déchet électroniqueavec les déchets usuels nonsoumis au tri sélectif. Se con-former aux ordonnances lo-cales sur les déchets munici-paux pour les dispositionsd’élimination et communi-quer avec le service à la cli-entèle pour des renseigne-ments sur les options d’élimi-nation responsable.

The following table applies only to the Ion Chef™ Instrument.

Conformitymark Description

Indicates conformity with safety requirements for Canada and U.S.A.

Indicates conformity with European Union requirements for safety andelectromagnetic compatibility.

Indicates conformity with Australian standards for electromagneticcompatibility.

Conformitysymbols

Appendix D SafetySymbols on this instrument D


Location of safety labels on this instrument

Safety information for instruments not manufactured by ThermoFisher Scientific

Some of the accessories provided as part of the instrument system are not designed orbuilt by Thermo Fisher Scientific. Consult the manufacturer's documentation for theinformation needed for the safe use of these products.

Instrument safety

CAUTION! Do not remove instrument protective covers. If you remove theprotective instrument panels or disable interlock devices, you may be exposedto serious hazards including, but not limited to, severe electrical shock, laserexposure, crushing, or chemical exposure.

CAUTION! Moving and Lifting Injury. Improper lifting can cause painful andpermanent back injury.

Things to consider before lifting or moving the instrument or accessories:· Depending on the weight, moving or lifting may require two or more

persons.· If you decide to lift or move the instrument after it has been installed, do notattempt to do so without the assistance of others, the use of appropriatemoving equipment, and proper lifting techniques.

· Ensure you have a secure, comfortable grip on the instrument or accessory.· Make sure that the path from where the object is to where it is being moved

is clear of obstructions.· Do not lift an object and twist your torso at the same time. Keep your spine

in a good neutral position while lifting with your legs.

General

Physical injury

Appendix D SafetyLocation of safety labels on this instrumentD


· Participants should coordinate lift and move intentions with each otherbefore lifting and carrying.

· For smaller packages, rather than lifting the object from the packing box,carefully tilt the box on its side and hold it stationary while someone elseslides the contents out of the box.

CAUTION! Moving Parts. Moving parts can crush, pinch and cut. Keep handsclear of moving parts while operating the instrument. Disconnect power beforeservicing.

WARNING! Ensure appropriate electrical supply. For safe operation of theinstrument:

· Plug the system into a properly grounded receptacle with adequate currentcapacity.

· Ensure the electrical supply is of suitable voltage.· Never operate the instrument with the ground disconnected. Grounding

continuity is required for safe operation of the instrument.

WARNING! Power Supply Line Cords. Use properly configured and approvedline cords for the power supply in your facility.

WARNING! Disconnecting Power. To fully disconnect power either detach orunplug the power cord, positioning the instrument such that the power cord isaccessible.

CAUTION! Cleaning and Decontamination. Use only the cleaning anddecontamination methods specified in the manufacturer's user documentation.It is the responsibility of the operator (or other responsible person) to ensurethe following requirements are met:

· No decontamination or cleaning agents are used that could cause aHAZARD as a result of a reaction with parts of the equipment or withmaterial contained in the equipment.

· The instrument is properly decontaminated a) if hazardous material isspilled onto or into the equipment, and/or b) prior to having the instrumentserviced at your facility or sending the instrument for repair, maintenance,trade-in, disposal, or termination of a loan (decontamination forms may berequested from customer service).

· Before using any cleaning or decontamination methods (except thoserecommended by the manufacturer), users should confirm with themanufacturer that the proposed method will not damage the equipment.

Electrical

Cleaning anddecontamination

Appendix D SafetyInstrument safety D


The Ion Chef™ System uses a UV lamp which emits light at 254 nm. Under normaloperating conditions, the UV lamp is powered on when performing the cleaningprotocol. Safety interlocks are used to ensure that the UV lamp is not powered whenthe door is open.

Safety and electromagnetic compatibility (EMC) standards

The instrument design and manufacture complies with the standards andrequirements for safety and electromagnetic compatibility as noted in the followingtable:

Reference Description

EU Directive2006/95/EC

European Union “Low Voltage Directive”

IEC 61010-1

EN 61010-1

UL 61010-1

CSA C22.2 No.61010-1

Safety requirements for electrical equipment for measurement,control, and laboratory use – Part 1: General requirements

IEC 61010-2-010

EN 61010-2-010

Safety requirements for electrical equipment for measurement,control and laboratory use – Part 2-010: Particular requirementsfor laboratory equipment for the heating of materials

IEC/EN 61010-2-020 Safety requirements for electrical equipment for measurement,control and laboratory use – Part 2-020: Particular requirementsfor laboratory centrifuges

IEC 61010-2-081

EN 61010-2-081

Safety requirements for electrical equipment for measurement,control and laboratory use – Part 2-081: Particular requirementsfor automatic and semi-automatic laboratory equipment foranalysis and other purposes


Directive 2004/108/EC European Union “EMC Directive”

FCC Part 15 U.S. Standard “Industrial, Scientific, and Medical Equipment”

AS/NZS 2064 Limits and Methods of Measurement of ElectromagneticDisturbance Characteristics of Industrial, Scientific, andMedical (ISM) Radiofrequency Equipment

ICES-001, Issue 3 Industrial, Scientific and Medical (ISM) Radio FrequencyGenerators

Ultraviolet (UV)Safety

Safety compliance

EMC

Appendix D SafetySafety and electromagnetic compatibility (EMC) standardsD



Directive 2012/19/EU European Union “WEEE Directive” – Waste electrical andelectronic equipment

Directive 2011/65/EU European Union “RoHS Directive” – Restriction of hazardoussubstances in electrical and electronic equipment

Environmentaldesign

Appendix D SafetySafety and electromagnetic compatibility (EMC) standards D


Instrument safety — Ion S5™/Ion S5™ XL System

For detailed information on Ion S5™ and Ion S5™ XL instrument safety symbols andalerts, safety and electromagnetic compatibility standards, and general instrumentsafety, see the Safety appendix of the Ion S5™ and Ion S5™ XL Instrument User Guide(Pub. No. MAN0010811), available at thermofisher.com.

Chemical safety

WARNING! GENERAL CHEMICAL HANDLING. To minimize hazards,ensure laboratory personnel read and practice the general safety guidelines forchemical usage, storage, and waste provided below. Consult the relevant SDSfor specific precautions and instructions:

· Read and understand the Safety Data Sheets (SDSs) provided by thechemical manufacturer before you store, handle, or work with any chemicalsor hazardous materials. To obtain SDSs, see the “Documentation andSupport” section in this document.

· Minimize contact with chemicals. Wear appropriate personal protectiveequipment when handling chemicals (for example, safety glasses, gloves, orprotective clothing).

· Minimize the inhalation of chemicals. Do not leave chemical containers open.Use only with adequate ventilation (for example, fume hood).

· Check regularly for chemical leaks or spills. If a leak or spill occurs, followthe manufacturer's cleanup procedures as recommended in the SDS.

· Handle chemical wastes in a fume hood.· Ensure use of primary and secondary waste containers. (A primary waste

container holds the immediate waste. A secondary container contains spillsor leaks from the primary container. Both containers must be compatiblewith the waste material and meet federal, state, and local requirements forcontainer storage.)

· After emptying a waste container, seal it with the cap provided.· Characterize (by analysis if necessary) the waste generated by the particular

applications, reagents, and substrates used in your laboratory.· Ensure that the waste is stored, transferred, transported, and disposed of

according to all local, state/provincial, and/or national regulations.· IMPORTANT! Radioactive or biohazardous materials may require special

handling, and disposal limitations may apply.

WARNING! HAZARDOUS WASTE (from instruments). Waste produced bythe instrument is potentially hazardous. Follow the guidelines noted in thepreceding General Chemical Handling warning.

Appendix D SafetyInstrument safety — Ion S5™/Ion S5™ XL SystemD


https://www.thermofisher.com

Biological hazard safety

WARNING! BIOHAZARD. Biological samples such as tissues, body fluids,infectious agents, and blood of humans and other animals have the potential totransmit infectious diseases. Conduct all work in properly equipped facilitieswith the appropriate safety equipment (for example, physical containmentdevices). Safety equipment can also include items for personal protection, suchas gloves, coats, gowns, shoe covers, boots, respirators, face shields, safetyglasses, or goggles. Individuals should be trained according to applicableregulatory and company/ institution requirements before working withpotentially biohazardous materials. Follow all applicable local, state/provincial,and/or national regulations. The following references provide generalguidelines when handling biological samples in laboratory environment.

· U.S. Department of Health and Human Services, Biosafety in Microbiologicaland Biomedical Laboratories (BMBL), 5th Edition, HHS Publication No. (CDC)21-1112, Revised December 2009; found at:www.cdc.gov/biosafety/publications/bmbl5/BMBL.pdf

· World Health Organization, Laboratory Biosafety Manual, 3rd Edition,WHO/CDS/CSR/LYO/2004.11; found at:www.who.int/csr/resources/publications/biosafety/Biosafety7.pdf

Appendix D SafetyBiological hazard safety D


http://www.cdc.gov/biosafety/publications/bmbl5/BMBL.pdf

http://www.who.int/csr/resources/publications/biosafety/Biosafety7.pdf

Documentation and support

Customer and technical support

Visit thermofisher.com/support for the latest in services and support, including:• Worldwide contact telephone numbers• Product support, including:

– Product FAQs– Software, patches, and updates– Training for many applications and instruments

• Order and web support• Product documentation, including:

– User guides, manuals, and protocols– Certificates of Analysis– Safety Data Sheets (SDSs; also known as MSDSs)

Note: For SDSs for reagents and chemicals from other manufacturers,contact the manufacturer.

Limited product warranty

Life Technologies Corporation and/or its affiliate(s) warrant their products as set forthin the Life Technologies' General Terms and Conditions of Sale found on LifeTechnologies' website at www.thermofisher.com/us/en/home/global/terms-and-conditions.html. If you have any questions, please contact LifeTechnologies at www.thermofisher.com/support.


http://thermofisher.com/support

http://www.thermofisher.com/us/en/home/global/terms-and-conditions.html

http://www.thermofisher.com/us/en/home/global/terms-and-conditions.html

http://www.thermofisher.com/support

For support visit thermofisher.com/support or email [email protected]

thermofisher.com

23 January 2017

http://thermofisher.com/support

mailto:[email protected]

http://thermofisher.com

Documents

Ion 540 Kit – Chef - Thermo Fisher Scientific · PDF fileNote: Ion 540™ Kit – Chef (Cat. No. A30011), for use when performing two sequencing runs per initialization, only supports