Establishment of high-sensitive multiplex typing systems for Y-STRs and its application to a very small amount of specimens

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In the preliminary step toward the present study, two novel multiplex typing systems for autosomal STR loci were constructed. These contributions were published in the journals. One of them was specially designed to have high sensitivity of the system. In order to select the loci with higher discriminating power, the haplotypes at 14 Y-STR loci were analyzed from two regional Japanese DNA samples (207 in Honshu and 87 in Okinawa) and 117 Thai samples using the 10-plex (DYS436, DYS439, DYS435, DYS19, DYS460, Y-GATA-H4, DYS391, DYS392, DYS438, DYS437) devised by NIST in the USA and the commercially released Y-PLEX6 kit (DYS393, DYS19, DYS389, DYS390, DYS391, DYS385) because of having two loci (DYS19 and DYS391) in common in both systems. Based on these studies, we completed to construct two kinds of high-sensitive multiplex systems for highly degraded DNA samples. One was for three loci (DYS392, DYS438 and DYS393) labeled with three-colored fluorescent dyes and another was for four loci (DYS19, DYS3891, DYS438 and DYS460) labeled with three-colored ones. The latter was especially useful because it made it possible to type the 4 loci from 50 pg of template DNA at usual numbers of PCR cycles (30 cycles) and from less amount of the template by increasing the number of cycles. Since the haplotype diversity was also calculated from 94 Japanese male samples, and the value was very high with 0.972, it was indicated this system could be very useful in forensic fields from the point of discriminating power as well. These results will be submitted elsewhere in near future. By this grant, our study to construct the multiplex system has been achieved to the goal.