Short tandem repeats on the Y chromosome (Y-STRs) are widely used in forensic laboratories for relationship testing, paternal lineage identification, complex kinship analysis, and in forensic cases such as sexual assault, missing person identification, disaster victim identification, and familial searching of suspects. The Microreader™ Y Prime Plus ID System (YPP) is a 6-dye, 38-locus commercially available Y-STR kit capable of direct amplification of samples and amplification of extracted DNA samples. It amplifies 37 Y-STR loci (29 loci from commonly used Yfiler® Plus PCR Amplification Kit & PowerPlex® Y23 System: DYS393, DYS570, DYS19, DYS392, DYS549, Y-GATA-H4, DYS460, DYS458, DYS481, DYS635, DYS448, DYS533, DYS449, DYS456, DYS389I, DYS390, DYS389II, DYS438, DYS391, DYS439, DYS437, DYS385a/b, DYS643, DYS518, DYS576, DYF387S1a/b, and DYS627) and 6 commonly used loci for the Y-STR database (DYS444, DYS447, DYS596, DYF404a/b, DYS527a/b, DYS557) as well as the Y-indel specifically designed for the Chinese population.
This study examined the validation of the reduction of the normal reaction volume of the Microreader™ YPP System from 25 μL to 5 μL to determine its efficiency in a smaller reaction system as a cost-effective measure. The reduced (5 μL) and full (25 μL) volumes methods were tested in triplicates by examining the analytical threshold, average peak heights, and the stutter ratio threshold. The reduced volume method’s analytical threshold was observed to be 107.96 RFU and 86.44 RFU for the full volume method. No significant difference was observed (at p-value > 0.05) in the average peak heights (p-value = 0.4505) as well as the stutter ratio threshold (p-value =0.2360) for both the reduced and full volume reaction methods.
This study demonstrates that the reduced volume reaction of the Microreader™ YPP System provides robust, reproducible, and reliable PCR amplification results that are comparable and similar to the original full reaction volume thereby promoting cost-effectiveness.
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