1Proprietary & ConfidentialThe world leader in serving scienceProprietary & Confidential

Philipp Habermeier

Sr. Key Account Manager HID

May 16, 2014

Higher discrimination power for human identification

– The new HID Ion AmpliSeq Identity and Ancestry

Panels for human identity applications.

2Proprietary & Confidential

Forensic DNA Analysis today

STR & CE – results in majority of samples

Cost effective & fast!

Accepted in court

Workflow validated & labs accredited

Enables DB‘s with up to 60% hit rate (e.g. UK!)Increased Efficiency

3Proprietary & Confidential

What is missing then – Where are the challenges?

Degraded samples

No / partial STR profiles

Difficult mixtures

Unknown sample origin

STR profile, but no DB hits

mtDNA

4Proprietary & Confidential

Use a new tool – Ion PGM™ System

Traditional Forensic Workflow:

Extend your workflow with the Ion PGM™ System

5Proprietary & Confidential

Use a new tool – Ion PGM™ System

6Proprietary & Confidential

Ion PGM™ Sequencer

Ion OneTouch™ 2 InstrumentsEmulsion PCR and Enrichment

Semiconductor Chips

Sequencing Chemistry• Natural nucleotides

• Natural enzymes

Sample Prep• Libraries

• Clonal beads

Torrent Server

The Ion PGM™ System

INSTRUMENTS REAGENTS DATA ANALYSIS

Ion 314™

Chip

Ion 318™

Chip

Ion 316™

Chip

7Proprietary & Confidential

Go from sample to results typically in 1-2 days!

9Proprietary & Confidential

HID-Ion AmpliSeqTM Identity Community Panel

For Research, Forensic or Paternity use only.

Not for diagnostic procedures.

10Proprietary & Confidential

For HID research use

99.71% concordant with published SNPs with high heterozygosity

and low Fst

Suitable for degraded DNA (Ave amplicon: ~136.9bp)

Recommended Input: ~1ng*

HID-Ion AmpliSeqTM Identity Community Panel

120 SNPs

30 Y -

SNPs

43 - Ken Kidd

SNPs

48 - SNPforID

SNPlex

* External collaborators (W. Parson) have seen acceptable sub-nanogram performance (≤500 pg)

11Proprietary & Confidential

HID SNP Genotyper Software

Input BED File

Pre

load

ed d

efa

ult p

ara

mete

rs b

ased o

n

Variant C

alle

r P

ara

mete

r settin

gs

12Proprietary & Confidential

HID SNP Genotyper Software - Output

Summary report

Export

output table

Easily

visualize

homozygous

and

heterozygous

genotypes

13Proprietary & Confidential

Comparing Genotypes

14Proprietary & Confidential14

Probability of Identity

85 unlinked autosomal SNPs from 1000 Genomes data, PI calculated from 1000 Genomes

11 loci

1 loci

7 loci

Amplicon size decreasing (e.g. for degraded DNA)

6E-35 – 1E-31

1E-11 – 1E-10

21 loci

The identity SNP panel has significantly lower

combined match probabilities than the standard

STR CE assay at all levels of degradation

15Proprietary & Confidential

HID-Ion AmpliSeq™ Ancestry Panel

For Research, Forensic or Paternity use only.

Not for diagnostic procedures.

16Proprietary & Confidential

Application: Subject exclusion and investigative leads that provides high discrimination between ancestral groups

165 autosomal markers (casework samples, investigative leads)

• 55 Ken Kidd (55 AIM set 1)

• average marker size of 130 nts

123 Seldin 2

• average marker size of 122 nts

16

HID-Ion AmpliSeq™ Ancestry Panel

1 Kidd, K. K. (2012). Better SNPs for Better Forensics: Ancestry, Phenotype, and Family Identification. Poster

2 Nassir, R., Kosoy, R., Tian, C., White, P. A., Butler, L. M., Silva, G., et al. (2009). An ancestry informative marker set for determining continental origin: validation and extension using human genome diversity panels. BMC Genetics, 10(1), 39.

Ken Kidd

Amplicons

Seldin

Size (nts) Ken Kidd

165 SNPs

Seldin

123

55

15

17Proprietary & Confidential17

SNP Source and Location

18Proprietary & Confidential18

Heat Map SNP Genotyper™

Ken Kidd 55 AIM SNPs, data from FROGdb

19Proprietary & Confidential

HID-Ion AmpliSeq™ Ancestry Panel

For 97% of autosomal SNPs to have at least 300x coverage,

target an average coverage of 594x.

At 97,890 reads per individual:

6 individuals per 314™ chip

30 individuals per 316™ chip

59 individuals per 318™ chip

19

Run recommendations by throughput

80% Loading, 60% Useable Reads

20Proprietary & Confidential

HID-Ion AmpliSeq™ Identity Panel

For 97% of autosomal SNPs to have at least 300x coverage,

target an average coverage of 738x. For 97% of Y-SNPs to

have at least 150x coverage, target an average coverage of 236x.

At 74,444 reads per individual:

8 individuals per 314™ chip

38 individuals per 316™ chip

77 individuals per 318™ chip

20

Run recommendations by throughput

80% Loading, 60% Useable Reads

21Proprietary & Confidential

Conclusion – NGS is capable to address these issues

Degraded samples

No / partial STR profiles

Difficult mixtures

Unknown sample origin

STR profile - but no DB hits

mtDNA

22Proprietary & Confidential

Mixture analysis tools

Body Fluid/Tissue identification

Additional externally visible phenotypes

Future applications

23Proprietary & Confidential

Want to learn more? Join our PGM workshops!

Fully

Booked!

24Proprietary & Confidential

Want to learn more? Join our PGM workshops!

Last few

remaining

spaces left!

To register please visit

https://learn.lifetechnologies.com/europe/humanidentification

25Proprietary & Confidential

For Research, Forensic or Paternity Use Only. Not for use in

diagnostic procedures.

When used for purposes other than Human Identification the

instruments and software modules cited are for Research Use

Only. Not for use in diagnostic procedures.

© 2014 Thermo Fisher Scientific Inc. All rights reserved. All

trademarks are the property of Thermo Fisher Scientific and its

subsidiaries unless otherwise specified.

Disclaimer

26Proprietary & Confidential

Visit our website at http://www.lifetechnologies.com/uk/en/home/ind

ustrial/human-identification/next-gen-sequencing-for-forensics.html

Please contact your local sales rep if you have any questions!