Genetic analysis for Y chromosome short tandem repeat haplotypes of Chinese Han population residing in the Ningxia province of China.

摘要

Whole-blood samples were obtained by venipuncture and collected into ethylenediaminetetraacetic acid (EDTA) tubes. Genomic DNA was extracted using the Chelex-100 protocol as described by Walsh et al. (1). Eleven Y-chromosomal short tandem repeats (STRs) loci (DYS19, DYS389I, DYS389II, DYS390, DYS391, DYS392, DYS393, DYS438, DYS439, and DYS385a,b) were performed in a fluorescence-based multiplex reaction using the Y-PLEX~(TM)12 kit (Reliagene Technologies Inc., New Orleans, LA). The amplification reactions of lCL in total contained 4.0 muL of 2.5 x Y-PLEX~(TM)12 Primer Mix, 0.20 muL of AmpliTaq Gold DNA Polymerase (Applied Biosystems, Foster City, CA) (5 UL), 3.8 muL of deionized water, and 2.0 muL of Genomic DNA (c. 1 ng). Thermal cycling conditions were conducted according to the manufacturer's kit protocols using Gene Amp PCR system 9700 (Applied Biosystems) (2). Detection and genotyping of allpolymerase chain reaction (PCR) products were accomplished using an ABI 3100 DNA Genetic Analyzer (Applied Biosystems). Allele designations were determined by comparison of the sample PCR fragments, together with the allelic ladders provided with the kit using GeneScan and Y12-Typer3100v 2.0 Macros (Reliagene Technologies Inc.).