2个miniSTR等位基因分型标准物的制备

目的采用分子克隆技术制备miniSTR D3S4529和D12ATA63基因座等位基因分型标准物,并评价其应用价值。方法用荧光引物对835份无关个体血卡样本进行扩增并分型检测,筛选2个基因座的等位基因片段,用分子克隆方法制备等位基因分型标准物,并对中国汉族群体进行遗传学调查。结果根据筛选出的等位基因片段制备出分型标准物,各等位基因峰形尖锐,无双肩峰,峰高基本一致,荧光值在4 000 RFU左右。中国汉族人群D3S4529、D12ATA63基因座分别检出7个、11个等位基因,杂合度分别为0.752、0.723,多态信息含量均为0.71。结论通过分子克隆法制备的miniSTR D3S4529和D12ATA63等位基因分型标准物,在法医学研究中具有较高的应用价值。

Preparation for standard allelic ladders of two miniSTR loci

Objective To construct two standard miniSTR（ D3S4529 and D12ATA63 ） allelic ladders by molecular cloning technology, and evaluate their application. Methods 835 unrelated individuals samples were amplified by fluorescence primer and analyzed by ABI 3130XL Genetic Analyzer to screen allele fragment. Standard allelic ladders were constructed by molecular cloning technique. To investigate the genetic polymorphism of D3S4529 and D12ATA63 loci in Han population. Results Standard allelic ladders were constructed by technology of molecular cloning. The alleles peak were sharp and balance. The fluorescent signal was about 4 000RFU. Investigation of the Han population genetics respectively showed 7 and 11 alleles. The heterozygosity observed were 0. 752 and 0. 723, respectively; and both polymorphism information component were 0. 71 for miniSTR D3S4529 and D12ATA63. Conclusion The two standard miniSTR（ D3S4529 and D12ATA63 ） allelic ladders were constructed by molecular cloning, which have high applicated value in forensic medicine research.