Validation of the Idseek® Omnistr™ Global Autosomal Str Profiling Kit, Reverse Complement Pcr as an Improved Tool/Method for Routine Massively Parallel Sequencing of Short Tandem Repeats

18 Pages Posted: 18 Jul 2024

See all articles by Kristiaan J. van der Gaag

Kristiaan J. van der Gaag

Netherlands Forensic Institute

Natalie E.C. Weiler

Netherlands Forensic Institute

Erik A.C. de Jong

affiliation not provided to SSRN

Jerry Hoogenboom

Netherlands Forensic Institute

Pieter van Oers

affiliation not provided to SSRN

Rick H. de Leeuw

affiliation not provided to SSRN

Elisabeth S.M. Graaf

Netherlands Forensic Institute

Thirsa Kraaijenbrink

affiliation not provided to SSRN

Joop Theelen

affiliation not provided to SSRN

Titia Sijen

Netherlands Forensic Institute

Abstract

Massively Parallel Sequencing (MPS) has gained interest in the forensic community over the past decade. Most of the published MPS methods focus on specialty applications intended for use in a limited number of samples with protocols that are relatively laborious. Recent developments using Reverse-Complement PCR enable an efficient MPS protocol suited for routine analysis of high numbers of samples. This method is implemented in the IDseek® OmniSTR™ Global Autosomal STR Profiling kit (Nimagen) for sequencing 28 of the most commonly used forensic autosomal STRs, one Y-chromosomal STR and Amelogenin.This study describes the validation of this kit and focuses on sensitivity, inhibitor tolerance, sequence variation detection and performance with mixtures up to 5 contributors. Results are compared to a Capillary Electrophoresis method (the PowerPlex® Fusion 6C system, Promega) and the first commercial forensic MPS kit (ForenSeq™ DNA Signature prep, Qiagen). Analysis settings in FDSTools are deduced and discussed, and an almost completely automated analysis is achieved. Using FDSTools noise correction, contributions in a mixture down to a level of 1.5% of the major allele of a marker can be detected.

Keywords: MPS, STR sequencing, IDseek OmniSTR, SE33, FDSTools, Mixture analysis, PCR inhibition, Reverse complement PCR

Suggested Citation

van der Gaag, Kristiaan J. and Weiler, Natalie E.C. and de Jong, Erik A.C. and Hoogenboom, Jerry and van Oers, Pieter and de Leeuw, Rick H. and Graaf, Elisabeth S.M. and Kraaijenbrink, Thirsa and Theelen, Joop and Sijen, Titia, Validation of the Idseek® Omnistr™ Global Autosomal Str Profiling Kit, Reverse Complement Pcr as an Improved Tool/Method for Routine Massively Parallel Sequencing of Short Tandem Repeats. Available at SSRN: https://ssrn.com/abstract=4898297 or http://dx.doi.org/10.2139/ssrn.4898297

Kristiaan J. Van der Gaag (Contact Author)

Netherlands Forensic Institute ( email )

Netherlands

Natalie E.C. Weiler

Netherlands Forensic Institute ( email )

Netherlands

Erik A.C. De Jong

affiliation not provided to SSRN ( email )

No Address Available

Jerry Hoogenboom

Netherlands Forensic Institute ( email )

Netherlands

Pieter Van Oers

affiliation not provided to SSRN ( email )

No Address Available

Rick H. De Leeuw

affiliation not provided to SSRN ( email )

No Address Available

Elisabeth S.M. Graaf

Netherlands Forensic Institute ( email )

Netherlands

Thirsa Kraaijenbrink

affiliation not provided to SSRN ( email )

No Address Available

Joop Theelen

affiliation not provided to SSRN ( email )

No Address Available

Titia Sijen

Netherlands Forensic Institute ( email )

Netherlands

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