The highest DoC for iSNP loci was for six loci (rs1109037 showing rs140313; rs192655; 14031320, rs4364205 and rs8037429) with a 140000-220,000 coverage depth. The rest of SNPs showed a DoC below were below 40,000. The AC) was calculated by UAS as a ratio of the number of reads for two alleles in heterozygous loci (Lower coverage/Higher Coverage). For SNP loci, ACR is equivalent to the ratio of reference allele coverage to reference and alternative allele coverage. The average ACR values obtained were 0.83 ± 0.069 for SNPs. Overall, the ACRs were quite even across the loci. Across, all samples, between 90 and 100% of the heterozygous loci, showed allelic balance ratios of ∼ 0.5. This study identified several novel iSNPs (Fig. 1). The novel variants were compared to the human genome reference sequence (GRCh38) (Supplementary Materials Table 1) [[8]Phillips C. Gettings K.B. King J.L. Ballard D. Bodner M. Borsuk L. Parson W. The devil’s in the detail”: release of an expanded, enhanced and dynamically revised forensic STR Sequence Guide.

Forensic Sci. Int. Genet. 2018; 34: 162-169

,[10]Young B. Faris T. Armogida L. A nomenclature for sequence-based forensic DNA analysis.

Forensic Sci. Int. Genet. 2019;

]. The data showed that most frequencies were above 0.5 indicating the forensic value of the markers. Autosomal STR (auSTR) and iSNP Random Mathc Probabilities (RMPs) for both length match probability and sequence match probabilities were calculated using STRait Razor software (Fig. 1A). For auSTRs, these values were 2.21 × 10-31 and 1.29 × 10-35. For iSNPs, these were 9.5 × 10-38 and 8.7 × 10-41. Loci rs2056277, rs722098, rs97118 and rs938283 showed the highest increases in RMP due to high heterozygosity in the Qatari population. Such data have not been reported earlier. The locus rs2056277 was previously reported by Poulsen et al. (2011) to be relatively uninformative for identity testing purposes. The iSNP full profile RMP was calculated to be 1.08 × 10-32. STRait Razor was used to generate the combined RMPs for auSTRs (Length-Based (LB) and Sequence-Based (SB) alleles) and iSNPs were 2.9 × 10-68 and 1.12 × 10-75.

Fig. 1(A) Bar chart showing the Random Match Probability (RMP) increase due to Sequence-Based Alleles. These were detected in Massively Parallel Sequencing of identity SNPs (The Combined Match Probability = 2.9 × 10-68 and 1.12 × 10-75). (B) Bar chart presenting the Identity SNP markers allele frequencies for the Qatari population. The samples were typed using ForenSeq™ DNA Signature kit and MPS analysis was done using Miseq-FGx. Frequency for most of the markers was above 0.4 showing the efficacy of these markers for individual identification.