Developmental validation of the IDseek® OmniSTR™ global autosomal STR profiling kit

Forensic science takes advantage of population variability in autosomal Short Tandem Repeat (STR) lengths to establish human identification. The most common method for DNA profiling by STR is based on PCR, where the highly polymorphic STR regions are amplified and analysed using Capillary Electrophoresis (CE) or Massively Parallel Sequencing (MPS). MPS determines not only the repeat length, but also the repeat structure and variations in the flanking regions, making this method superior in discriminatory power compared to CE. Reverse Complement PCR (RC-PCR) is a novel, more sophisticated PCR based MPS library preparation method combining indexing and PCR amplification in a single closed-tube reaction. In this document we describe the complete developmental validation of the IDseek® OmniSTR™ kit, an RC-PCR based MPS library preparation kit. The developed IDseek® OmniSTR™ kit contains 28 autosomal STR targets, one Y-chromosomal STR and the Amelogenin gene covering all relevant STR core loci from the USA, EU, UK and Interpol. •RC-PCR is an MPS library prep method combining indexing and amplification in a single tube reaction.•The OmniSTR™ kit displays uniform amplification with a heterozygosity ≥ 0.6 in 97.80 % of samples.•OmniSTR™ shows high sensitivity with allelic recovery > 84 % for as little as 15.60 pg DNA input.•In 1:20 mixtures, 72.87 % of the unshared minor contributor alleles are still detected.•Proven tolerance for common co-extracted PCR inhibitors, such as Humic and Tannic acid and Hematin.