Excluding the primate species, no peaks were detectable above 50 RFU in either PowerPlex® ESI 17 Fast or ESX 17 Fast for any of the domestic animal or microbial samples except
for A. lwoffi which showed a XAV-939 in vivo peak at 292–293 bases with a height of 94–108 RFU in the green dye channel (D10S1248) of PowerPlex® ESI 17 Fast and also at 89–90 bases with a height of 116–129 RFU in the green dye channel (D10S1248) of PowerPlex® ESX 17 Fast. In both cases the peaks migrated on-ladder as an 11 allele. A. lwoffi is part of the normal oropharynx and skin flora of about 25% of individuals [27] with a genome size of 3.48 Mb. At 10 ng DNA in an amplification reaction, this equates to 2.6 million genome copies which suggests a low level cross hybridization to give a peak of the height seen. Primates show the most amplification peaks with fewest being seen with macaque, followed
by gorilla and orangutan with a comparable number of peaks and the greatest number with chimpanzee (Supplemental Fig. 18). The results from the 20 mock crime stain samples amplified with the PowerPlex® ESI Fast Systems were either the same or better than the equivalent AmpFlSTR® SGM Plus® results in terms of number of alleles recovered (Supplemental Table 6). Full profiles were concordant Everolimus with the AmpFlSTR® SGM Plus® profile of the major donor. In general the 44 mock crime stain samples amplified with the PowerPlex® ESX Fast Systems generated allele calls that were concordant with those obtained with the Investigator®
ESSplex Plus Kit with recovery of similar numbers of alleles (Supplemental Table 7). However, a few samples had apparent discordant allele calls. In the case of samples 13-031518R-01-1, why 13-031880R-01-1, and 13-031881R-01-1, the PowerPlex® ESX Fast Systems called a 16.3 allele at D1S1656. This was labelled off-ladder with the Investigator® ESSplex Plus Kit due to the allele migrating just to the right edge of the 16.3 allele bin position. In addition, in sample 13-031881R-01-1 there was an apparent discordance at D1S1656 with the Investigator® ESSplex Plus Kit calling this as an OL, 17.3 whereas PowerPlex® ESX 17 Fast genotyped this sample as 16.3, 18.3. This sample was a low level mixture (PowerPlex® ESX 17 Fast profile showed three alleles at the SE33 locus) and gave a partial profile for the major contributor with Investigator® ESSplex Plus and a full profile for the major contributor with PowerPlex® ESX 17 Fast. However, amplification of the major contributor to this mixture with PowerPlex® ESI 17 Fast and ESX 17 Fast (primer pairs for D1S1656 being different between these two multiplexes) gave a genotype of 16.3, 18.3 with both kits as well as with the Investigator® ESSplex Plus Kit (Supplemental Fig. 19). Thus, the discordance seen with this casework sample appears to be due to this being a low level mixture with the 17.3 allele possibly being due to the second minor contributor in this mixed casework sample.