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HID University Seminar Series Intro & Evolution of Forensic DNA
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Future Trends in Forensic DNA Technology Seminar Series
Introduction Human Identification Life Technologies
8/16/2012 2
GROUNDED in Science INSPIRED by Scientists DRIVEN by Innovation
1 Company / 8 Trusted Brands / 600,000+ Citations
8/16/2012 3
Quick Facts
8/16/2012 4
Media Recognition
Ranked #9 in 2012
Red Dot 2012 Award for Excellence In Product Design
Ranked #15 in 2012
Ranked Among “The 50 Most Innovative
Companies in 2012”
Austin Plant Awarded Best Plant in 2012
318 Chip Awarded for Design & Engineering
In Embedded Technologies
8/16/2012 5
Ion Semiconductor Sequencing Rapid, Benchtop Sequencing for All
8/16/2012 6
This is Why We’re Here
“When you are in prison, and you are told that you’ve got to be
there for the rest of your life - for something you didn’t do –
DNA testing said otherwise”
Herman Atkins, Exonerated February 2000
“the new super toxic E. Coli strain was decoded in just two hours – a marvel to scientists”
June 2011
“Whole-genome sequencing has enabled doctors to provide the
Beery twins with a simple, highly effective treatment for a
rare condition”
June 2011
Freedom for the Innocent Solutions to Epidemics Life Changing Medicine
8/16/2012 7
This is Why We’re Here
Reported rapes have fallen to the lowest level in 20 years as DNA evidence helps send more rapists to prison
USA Today, October 7, 2009
8/16/2012 8
Combating Human Trafficking A Major Global Problem
An estimated 600,000 to 820,000 men, women, and children are trafficked across international borders each year, approximately 80 percent are women and girls and up to 50 percent are minors – US State Department
The human trafficking phenomenon affects virtually every country, including the United States – Hillary Clinton, US Secretary of State
12.3 million adults and children in forced labor, bonded labor, and commercial sexual servitude at any given time…. estimated 56 percent of all forced labor victims are women and girls – International Labor Organization (United Nations)
8/16/2012 9
Use of DNA Testing to Prevent Human Trafficking
In January 2011, more than 70 people including 25 children from Haiti arrived at an airport in Bolivia from Lima. Visa problems stopped them on their way to Brazil or Argentina
Bolivian Police suspicions opened an investigation. The Bolivia Forensic Research Institute, a DNAProkids laboratory, performed DNA testing using Bode swabs and the Identifiler® Direct kit
All the children were being trafficked: 13 were returned to their families in Haiti who were searching for them
Route Taken to Bolivia from Haiti
8/16/2012 10
8/16/2012 11
Customer Product Communication System
A web-based system that ensures the timely delivery, and real-time reporting, of important technical product information to appointed contacts at each North America customer site. Launched in North America October 2011
Communication about critical product quality issues affecting specific HID customers: product retirement, product discontinuation, new software version or patch release, user bulletin, and non-critical technical updates
Not a marketing channel!
To sign up, go to: http://marketing.appliedbiosystems.com/mk/get/ HID_Product_Notification_Sign_Up
Minor Manufacturing Changes to the AmpFSTR® PCR Amplification Kits
New Vendor Qualification Program
8/16/2012 13
Minor Modifications to Several AmpFSTR® Kits
Manufacturing of the AmpliTaq Gold® enzyme and 10X PCR Buffer II components transitioning from Roche to Life Technologies
− Both components in the following kits: Profiler®, Profiler Plus®, Profiler Plus® ID, SGM Plus®, COfiler®, Identifiler®, Yfiler® & MiniFiler™ Kits
− The buffer component only in the Identifiler® Direct Kit (this kit already contains the AmpliTaq Gold® enzyme manufactured by Life Technologies)
− Manufacturing of both components by Life Technologies will be conducted according to the same specifications used previously by Roche.
− All other kits already contain both the buffer and enzyme components manufactured by Life Technologies
8/16/2012 14
Minor Modifications to Several AmpFSTR® Kits
Our internal studies show the kits containing the components manufactured by Roche and Life Technologies to be interchangeable − The buffer and enzyme components manufactured by Life Technologies
have already been tested rigorously during developmental validation of the Identifiler® Plus, NGM™ and NGM SElect™ Kits
User Guides and/or Product Inserts for each of the affected kits will be updated to include our internal developmental validation studies demonstrating no substantive difference in performance compared to previous versions
A full revalidation should not be necessary; however, laboratories should determine their own implementation requirements
8/16/2012 15
Minor Modifications to Several AmpFSTR® Kits
Transition Process − Expected to commence in September 2012 − New and current versions will be available simultaneously for approximately 3
months to allow customers to transition smoothly − All kits will maintain the same kit names and part numbers
> The first batch of kits containing buffer and enzyme components manufactured by Life Technologies can be distinguished from earlier versions by the letters “NF” after the lot number
Why are we doing it? − To exercise greater control over the quality of key raw materials − To prevent formulation or QC issues which affect kit performance − To better maintain reagent supply
8/16/2012 16
Forensic News Spring 2012 Edition
− www.appliedbiosystems.com/forensicnews − The latest information on human identification applications,
product updates, unique laboratory applications and new technologies
Includes articles on: − Tools for Improved DNA Extraction Efficiency
− Expanding the Capabilities of Direct Amplification
− Sample Collection Solutions from Life Technologies
− Coming Soon...The World's Most Powerful STR Kit
− Identifiler® Plus Kit Increases Success Rate of DNA Typing from Challenging Casework Samples by as much as 30%
− MiniFiler™ Applications to Human Identification and Forensic Casework Analysis
− Case Study: Y Testing at Court—AmpFSTR® Yfiler® Kit Drives the Success of Y Chromosomal Analysis in Forensic Genetics
− Considerations for the Evaluation of Plus Stutter for AmpFSTR® PCR Amplification Kits in Human Identification Laboratories
8/16/2012 17
ACTIVATE (Touch)
AIM (Autoscan)
RESULTS (COA info)
New Life Tech App – Technical Document App “Coming Soon” to an App Store near you
Sample - Search for COA
8/16/2012 18
Thank You!! © 2012 Life Technologies Corporation. All rights reserved. The trademarks mentioned herein are the property of Life Technologies Corporation or their respective owners.
Human Identification, Life Technologies
Evolution of Forensic DNA
8/16/2012 20
A look back… 10 years ago
9/11 WTC Attacks
CSI only in 2nd Season, but surging in popularity
8/16/2012 21
A look back… 10 years ago
Fragmented, Inefficient Forensic DNA Workflow − Manual Processes − Quantiblot − GeneScan®/Genotyper® Software
Minimal Use of Y-STRs
Limited options for analyzing heavily degraded samples
Global DNA Databases in their infancy (except for the UK)
8/16/2012 22
Common beliefs I encountered…
“Automation cannot work for forensic DNA casework.”
“Manual Organic Extraction is the only trusted method for casework.”
“Automated data analysis with advanced software (e.g. expert systems) is impossible.”
“Analysis of lesser felonies i.e. property crimes is not practical.”
8/16/2012 23
~1 in 1018 (quintillion)
Disc
rimin
atio
n Po
wer
Increased Discrimination and Sensitivity/Robustness
Bone sample amplified using the NGM™ Kit
Bone sample amplified using the Identifiler® Kit
Mea
n Pe
ak H
eigh
t (RF
U)
8/16/2012 24
Today Increased Integration and Efficiency…….
Increased Efficiency
Ease of Use Success with Difficult Samples
Simplified Interpretation
Higher Throughput
Streamlined Workflow
8/16/2012 25
Streamlined Workflows for Single Source Samples
NGM SElect™ Express Kit
Identifiler® Direct Kit
Prep-n-Go™ Buffer
8/16/2012 26
Total Time to Result = ~10-58 hrs
Sample Preparation
Quantitation/ Normalization Amplification Detection Analysis
Time to Result
(hrs/per 6-16 samples) 1-48 2 3.5 0.75 2-3
Dependent on sample type and method
Improved Efficiency for Casework Samples
8/16/2012 27
Total Time to Result = ~10-58 hrs
Sample Preparation
Quantitation/ Normalization Amplification Detection Analysis
Time to Result
(hrs/per 6-16 samples) 1-48 2 3.5 0.75 2-3 Time to Result
(hrs/per 13 samples) 1-3 2 1.5 0.55 1-2
Total Time to Result = ~6-9 hrs
Improved Efficiency for Casework Samples
8/16/2012 28
Evolution of STR Technology
1996 2012
Triplexes
Green I
Blue
Profiler Plus®
Profiler®
Profiler Plus® ID
COfiler®
SGM Plus®
Identifiler®
SEfiler™
Larger Multiplexes
Yfiler®
SEfiler™ Plus
MiniFiler®
Enhanced Capability
NGM™
NGM SElect™
NGM SElect™ Express
Identifiler® Plus
Identifiler® Direct
Optimized for Application
Expanded “Global”
Multiplexes
AmpFSTR® PCR Amplification Kits
A comprehensive product portfolio, developed, validated and manufactured according to stringent performance standards specifically for use on forensic samples
8/16/2012 29
UNITED STATES LEGISLATIVE WAVE
ALL CONVICTED OFFENDER LEGISLATION 1999 – 5 states 2003 – 27 states 2008 – 42 states 2011 – 50 states
ARRESTEE LEGISLATION 1999 – 1 state 2006 – 7 states 2011 – 26 states
8/16/2012 30
THE WORLD 44 COUNTRIES, OVER 35 MILLION OFFENDER SAMPLES
ADOPTS DATABASING Fully implemented programs Legislation passed, programs not yet implemented
Australia Austria Barbados Belarus Belgium Brazil Canada Chile China Croatia Cyprus Denmark
Estonia Finland France Germany Hong Kong Hungary Iceland Israel Japan Jordan Kuwait Latvia
Netherlands New Zealand Macedonia Norway Panama Poland Portugal Qatar Russia Slovenia Slovakia Singapore
South Korea Spain Sweden Switzerland Taiwan United Arab Emirates United Kingdom United States
8/16/2012 31
Prüm Treaty DNA Implementation Status 11-11-11
Progress Towards Cross-Border Data Exchange
8/16/2012 32
Global Databases: Locus Overview
Locus Europe CODIS CODIS Proposed
TPOX X* X
CSF1PO X* X*
D5S818 X* X*
D7S820 X* X*
D13S317 X* X*
FGA X* X* X*
vWA X* X* X*
D3S1358 X* X* X*
D8S1179 X* X* X*
D18S51 X* X* X*
D21S11 X* X* X*
TH01 X* X* X*
D16S539 X X* X*
Locus Europe CODIS CODIS Proposed
D2S1338 X X X*
D19S433 X X X*
D12S391 X* X*
D1S1656 X* X*
D2S441 X* X*
D10S1248 X* X*
D22S1045 X* X
SE33 X X
DYS391 X*
* Required loci
Loci in bold contained in either the Identifiler® or NGM SElect™ Kits
8/16/2012 33
The World Is Positioning For Explosive Growth of DNA Databases
Dr. Reddy, Laboratory Director CDFD, India
8/16/2012 34
INDIA INDONESIA BRAZIL (LEG. PASSED) PAKISTAN NIGERIA RUSSIA (LEG. PASSED) MEXICO VIETNAM TURKEY THAILAND ITALY (LEG. PASSED) SOUTH AFRICA
COLOMBIA ARGENTINA KENYA PERU SAUDI ARABIA MALAYSIA (LEG. PASSED)
“Currently there are about 35 trained analysts for DNA profiling [in all of India]. The country will need about 1,100 analysts [over the next 10 years].”
The World Is Positioning For Explosive Growth of DNA Databases
8/16/2012 35
Increasing Casework Backlogs U.S. Casework Backlogs
Source: NIJ Special Report “Making Sense out of DNA Backlogs,
2010 – Myths vs. Reality” www.ojp.usdoj.gov/nij
Backlogs increasing with Increased submissions
Samples increasing in complexity
Shortage of trained analysts
Validation and implementation barriers
To help address these and other challenges, we want to provide more than just core technologies
8/16/2012 36
HID Service Solutions HID Professional Services Program (HPS)
− Delivery of internal validation programs and/or performance checks on any Applied Biosystems HID product
− Robotic Validation/Implementation Support
“HID University” Training Programs − Comprehensive suite of training courses designed to provide continuing education
to forensic DNA analysts
LIFE Center for Forensic Excellence at UNTHSC − Helps developing nations to rapidly establish world-class database laboratories
through a unique certificate training program
Advisory Consulting
Complete Laboratory Integration and Validation
Analyst Training Programs
On-site Support and Training
8/16/2012 37
Mitochondrial DNA Sequencing
SNP Genotyping Paternity, Mixtures, Missing
Persons, DVI
Phenotypic SNPs for Investigative Leads
Eye Color
Hair Color
Missing Persons, DVI Facial Reconstruction
Looking ahead… Potential PGM™ Human Identification Applications
8/16/2012 38
21st Century Crime Fighting...
DNA profiles of all felony arrestees loaded to an investigative database
8/16/2012 39
21st Century Crime Fighting...
DNA used to investigate all types of evidence
8/16/2012 40
21st Century Crime Fighting...
Data shared globally across national databases
8/16/2012 41
21st Century Crime Fighting...
Data shared globally across national databases
Rapid turnaround time of DNA evidence
8/16/2012 42
Our Role Enable Forensic Labs to Do More with Less
• Increase efficiency with next-gen products and workflow integration (more info, less time)
• Improve performance on difficult and compromised samples (more info, less sample)
• Be a true systems integration partner (leveraging HID Professional Services)
• Enable use of latest technologies for forensic and biometric applications
8/16/2012 43
Thank You
© 2012 Life Technologies Corporation. All rights reserved. The trademarks mentioned herein are the property of Life Technologies Corporation or their respective owners
For Forensic & Paternity Use Only
The Evolution of Direct Amplification: From Sample to Result in 2 Hours*
Human Identification, Life Technologies
* Typical workflow for 24 samples
8/16/2012 45
Traditional Single Source Sample Workflow
Time to Result
(hrs/per 24 samples) ~1-3.5 ~0-2 ~4 ~1.5 ~1
Total Time to Result = ~7-12 hrs
Collection Sample Preparation
Quantitation/ Normalization Amplification Detection Analysis
8/16/2012 46
Development of Direct Amplification on Treated Paper Substrates
8/16/2012 47
Evaluation of Direct Amp Capability of the Identifiler® Kit
Control DNA 007 25 μL Reaction Volume
1.2 mm Blood on FTA® Disc
No full profiles under direct amp conditions
8/16/2012 48
Effect of Reducing Disc Size on Identifiler® Kit Direct Amp Performance
0.5 mm blood on FTA® disc
1.2 mm blood on FTA® disc
Control DNA 007 (1 ng)
0.75 mm blood on FTA® disc
But: − No automated
0.5 mm punch head options
− 0.5 mm disc size too small for buccal samples
− 0.5 mm discs very hard to handle
8/16/2012 49
Identifiler® Kit Direct Amp Performance Evaluation
Possible explanations for poor performance − Insufficient DNA?
− Excess PCR inhibitors?
0.75 mm disc ~ 22.08 ng DNA
0.5 mm disc ~ 9.81 ng DNA
1.2 mm disc ~ 56.52 ng DNA Height = 1mm
1 μL blood ~ 50 ng DNA
Sufficient DNA still available Inhibitor concentration reduced by >5-fold
0.5 mm disc ~ 9.81 ng DNA
8/16/2012 50
Optimization of Reaction Buffer for 1.2 mm Discs
Component 1
Response Component 2
Use of Design of Experiments (DOE) Approach
8/16/2012 51
Optimization of Identifiler® Direct Kit Master Mix Buccal sample on 1.2 mm FTA® disc before DOE
8/16/2012 52
Optimization of Identifiler® Direct Kit Master Mix Buccal sample on 1.2 mm FTA® disc after DOE
8/16/2012 53
− 3 x liquid blood samples − 80 μL of blood onto FTA® Classic (passive diffusion) − Sample 1.2 mm discs from the center to the edge of the blood stain − Perform replicates for each position
Effect of Punch Position on Sample Peak Heights
1 3 2 4 5
8/16/2012 54
Direct Amplification from Treated Paper
FTA® Classic Card
EasiCollect® System
Identifiler® Direct Kit
Identifiler® Kit
Exam
ple
1 Ex
ampl
e 2
Identifiler® Direct Kit
Identifiler® Kit
Blood on FTA® samples
8/16/2012 55
Identifiler® Direct Kit Validation: External Test Site Results
Sample Type
PCR Success Rate Interpretation Success Rate Number of
Samples Tested Range of Success rates
Mean Success Rate
Range of Success rates
Mean Success Rate
VTS Study
Blood 99.4 % 99.4 % 95.7 – 98.8% 97.3 % 414
Buccal 91.8 – 99.4% 97.1 % 84.2 – 95.5% 90.9 % 653
CTS Study
Blood 100% 100 % 98.8 – 100% 99.8 % 437
Buccal 98.7 – 100% 99.0 % 91.7 – 100% 94.7 % 703
1st Pass Success Rate Definition − All profile peaks higher than specified threshold − Off scale peaks produce no artifacts which interfere with profile
interpretation > OL-labelled pull-up peaks <20% of highest peak of the marker
> No split, double called peaks
> Stutter peaks < 20% of marker or < marker stutter cut off, whichever is higher
8/16/2012 56
Expansion of the Direct Amplification Workflow to Non-FTA Substrates
Untreated Paper
8/16/2012 57
Identifiler® Direct Kit development and optimization performed on FTA® substrates only
Laboratories using untreated paper or swab substrates and/or alternative marker sets looking to recognise time and cost savings of the Direct Amplification workflow
Expansion of the Direct Amplification Workflow to Non-FTA® Substrates
Goal: Enable direct amplification of non-FTA® buccal samples
Minimal additional workflow steps
High quality, well balanced profiles
No introduction of artifacts
Prep-n-Go™
Buffer
8/16/2012 58
Direct Amp Workflow: Untreated Paper Substrates
Collect Samples on Untreated Paper
One New Step
Electrophorese
Amplify
Punch 1.2 mm disc
Add PCR reagents
Add 2 µL Prep-n-Go™ Buffer Untreated paper
protocol development
performed on Bode Buccal Collector™
8/16/2012 59
Identifiler® Direct Kit Amplification of Bode Buccal DNA Collector™ Samples lysed with Prep-n-Go™ Buffer
2 different individuals amplified for 26 PCR cycles Well-balanced profiles within each dye color
8/16/2012 60
Performance Study Results Summary
Test Site Number of
Samples Tested
PCR Cycle
Number
CE Platform
PCR Success Rate Interpretation Success Rate
Number of Full Profiles
First Pass Success Rate
Number of Full Profiles
First Pass Success Rate
1 82* 27 3130xl 81/82 98.8% 78/82 95.1%
2 80 26 3130xl 78/80 97.5% 78/80 97.5%
3 84* 26 3130xl 83/84 98.8% 79/84 94.0%
4 84* 26 3130xl 80/81 98.8% 76/81 93.8%
5 84* 26 3730 83/84 98.8% 83/84 98.8%
6 84 26 3500 82/84 97.6% 74/84 88.1%
Life Technologies 84 26 3130xl 82/84 97.6% 79/84 94.0%
Total 582 569/582 97.8% 547/582 94.0%
* Used real offender samples
8/16/2012 61
Performance Study: Bode Buccal DNA Collector™ Samples lysed in Prep-n-Go™ Buffer
Peak Heights Intracolor Balance
All amplifications performed using the Identifiler® Direct Kit
8/16/2012 62
Identifiler® Direct Kit w/ Prep-n-Go™ Buffer Identifiler® Direct Kit w/Other Buffer
1200 1200
Lysis Buffer Performance Comparison
No allelic drop-out using Prep-n-Go™ Buffer
8/16/2012 63
Expansion of the Direct Amplification Workflow to Non-FTA Substrates
Buccal Swabs
8/16/2012 64
Pros − Easy to use − Inexpensive − Multiple types available
Buccal Swabs
Cotton Swab
Foam Swab
T-Swab
Flocked Swab
Omni Swab
8/16/2012 65
Cons − High performance variability
among swab types − Donor variation − Sample collection techniques
and storage conditions − Limited opportunities for
automation increasing labor requirements and the risk of contamination
Buccal Swabs
Donor Mouth Conditions Collection Technique
Long-Term Storage Swab Age
Drying & Transport Conditions
8/16/2012 66
Flocked Swab Swab Structure Comparison
Cotton Swab
2 km microfiber 6 m microfiber
Sample stays entrapped in the
fiber wand
Sample is released quickly and in
higher amounts
8/16/2012 67
4N6FLOQSwabs™ by Copan
Distributed by FLOQSwabs™ consist of short
Nylon® fibers that are arranged in a perpendicular fashion
Excellent recovery of DNA
Maximum efficiency in collection capacity
Available with different anatomical and ergonomic designs
Certified free of Human DNA, Dnase and RNase
ETO-treated (Ethylene Oxide)
8/16/2012 68
Direct Amp Workflow: Identifiler® Direct Kit
Collect Samples on Buccal Swab
Lyse swab in 400 μL Prep-n-Go™ Buffer
Add PCR Reagents
Transfer Lysate
New steps
Electrophorese
Amplify
Incubate in oven adaptor at 99°C for 20 minutes
8/16/2012 69
Performance of Aged Swabs: Identifiler® Direct Kit & Prep-n-Go™ Buffer
4N6FLOQSwabs™
Puritan Swabs
Omni Swabs
Mean time between collection and analysis: 90 days; Amplification: 26 cycles
8/16/2012 70
Expansion of the Direct Amplification Workflow to Other STR Marker Sets
8/16/2012 71
Direct Amplification with the NGM™ & NGM SElect™ Kits NGM SElect™ Kit NGM™ Kit
Amplification of buccal samples on Bode Buccal Collector™ treated with Prep-n-Go™ Buffer
8/16/2012 72
Utilizes the same primer sequences as the NGM™ and NGM SElect™ Kits
Includes a new master mix optimised specifically to support direct amplification of swab and treated/untreated paper substrates
Delivers rapid cycling times through the introduction of a new fast-capable enzyme (< 1 hr)
May be amplified using the Veriti® 96-Well or 9700 (silver or gold-plated silver block) thermal cyclers − Veriti® 96-Well Thermal Cycler (standard) now supported for use with all
existing AmpFSTR® kits
Direct Amplification for the Expanded ESSL
AmpFSTR® NGM SElect™ Express Kit
8/16/2012 73
Direct Amp Workflow: NGM SElect™ Express Kit
Collect Samples on Buccal Swab
Lyse swab in 400 μL Prep-n-Go™ Buffer
Add PCR Reagents
Transfer Lysate
New steps
Electrophorese
Amplify
Heated lysis optional
depending upon swab age/type
8/16/2012 74
AmpFSTR® NGM SElect™ Express Kit Example Profiles: Treated Paper
Blood on FTA® Classic Card
Buccal on Copan NUCLEIC-CARD™
8/16/2012 75
AmpFSTR® NGM SElect™ Express Kit Example Profiles: Untreated Paper
Blood on 903 Paper
Buccal on Bode Buccal Collector™
8/16/2012 76
AmpFSTR® NGM SElect™ Express Kit Example Profiles: Swabs
Buccal on Copan Flocked Swab
Buccal on Whatman Omniswab
8/16/2012 77
Test Site
Sample Type
Cycle #
CE Platform N
Profile Assessment
Partial Profile
No
Prof
ile
Off
Scal
e Pr
ofile
s
First Pass Success Rate
50 RFU
175 RFU
450 RFU
50 RFU 175 RFU 450 RFU
# % # % # %
Repr
oduc
ibili
ty 3 Copan 25 3500xL 60 1 1 1 1 1 58 97 58 97 58 97
4 Copan (2 weeks) 26 3100 60 0 4 N/A 1 37 58 97 58 97 NA NA
5 Copan (Fresh) 25 3130xl 60 0 0 N/A 0 8 60 100 60 100 NA NA
Perf
orm
ance
3 Omniswab 27 3500xL 40 1 2 3 1 0 37 93 36 90 35 88
4 Prionics (2 months) 26 3100 40 0 2 N/A 1 31 39 98 39 98 NA NA
5 Omniswab 26 3130xl 42 0 1 N/A 0 7 42 100 42 100 NA NA
Test Site Results
8/16/2012 78
8/16/2012 79
Factors Influencing Direct Amplification Results
Choice of kit
Choice of sample type/substrate Sample Transfer Efficiency
Punch Size and Position
Cycle Number
Data Analysis Parameters
8/16/2012 80
Maximising Direct Amplification Results
Swab Substrate Handling − Ensure swabs are fully dried before storing − Ensure swabs are stored correctly to prevent excessive degradation over
time
Swab Lysate Handling − If using a 96-well deep well plate for lysis, remove lysate from swab heads
when aliquotting for storage and discard the plate containing the swab heads to reduce contamination risk
− Avoid taking cell debris or precipitation from the bottom of the lysate tube when transferring swab lysate to amplification or storage plates/tubes
− Follow the kit-specific instructions for lysis and amplification > Heated lysis may improve performance dependent upon direct amplification kit, swab
age and type
8/16/2012 81
Maximising Direct Amplification Results
Thermal Cycling Platform − Life Technologies kits optimised for use on the 9700 with silver or gold-
plated silver block and the Veriti® 96-Well thermal cycler only > Not supported for use on the 9700 with Aluminium block or
> Not supported for use on the Veriti 96-well Fast thermal cycler
Choice of Cycle Number − Choose a cycle number that prevents allele drop-out and minimizes off-
scale alleles − Use of elevated cycle numbers may cause presence of artifacts
Optimisation of Software Analysis Settings − Use of settings appropriate to single source samples will reduce editing
requirements and facilitate expert system analysis
8/16/2012 82
Optimising Data Analysis Parameters for Single-Source Samples
Use of a global cut-off filter can reduce significantly the amount of editing required for single-source sample data
Peak Quality settings may also be adjusted to better reflect the characteristics of single-source samples
8/16/2012 83
Optimized Parameters = More Efficient Analysis U
nopt
imize
d Optim
ized
8/16/2012 84
Time to Result
(hrs/per 24 samples) ~1-3.5 ~0-2 ~4 ~1.5 ~1
Maximized Efficiency for Single Source Sample Processing
Total Time to Result = ~7-12 hrs
Collection Extraction Quantitation/ Normalization Amplification Detection Analysis
8/16/2012 85
Time to Result
(hrs/per 24 samples) ~1-3.5 ~0-2 ~4 ~1.5 ~1
Maximized Efficiency for Single Source Sample Processing
Time to Result
(hrs/per 24 samples) 0 0 0.75 0.55 0.1
Total Time to Result = ~7-12 hrs
Collection Amplification Detection Analysis
Total Time to Result = ~2 hrs
8/16/2012 86
*Kits currently in development
8/16/2012 87
Thank You © 2012 Life Technologies Corporation. All rights reserved. The trademarks mentioned herein are the property of Life Technologies Corporation or their respective owners
For Forensic or Paternity Use Only.