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目的对300℃焚烧后成人股骨样本进行9个miniSTR(D20S1082、D6S474、D12ATA63、D9S1122、D2S1776、D1S1627、D3S4529、D2S441、Amelogenin)基因座的检测与分型。方法样本为8根经300℃焚烧后的成人股骨,用改良酚-氯仿法提取烧骨DNA,在Mastercylcerpro梯度PCR仪上对9个miniSTR基因座分别进行扩增,3130基因分型仪检测并收集电泳结果,GeneMarkerV2.2.0软件计算扩增产物片段相对大小以及进行样本基因型分型。结果8根烧骨样本均能够提取到DNA,浓度平均值为25ng/μL,D260/D280值在1.7~1.9之间。9个miniSTR基因座在样本中的检出率在78%~100%之间,分型图谱较清晰,个别样本出现额外带。结论本文9个miniSTR基因座分型检测的方法,可用于对烧骨捡材的DNA分型检验。
Objective To detect and classify nine miniSTR (D20S1082, D6S474, D12ATA63, D9S1122, D2S1776, D1S1627, D3S4529, D2S441, Amelogenin) loci in adult femurs after incineration at 300 ℃. Methods Eight samples of adult femurs incinerated at 300 ℃ were collected. The bone DNA was extracted by modified phenol - chloroform method. The 9 miniSTR loci were amplified by Mastercylcer ® software and sequenced by 3130 genotyping instrument And collect electrophoresis results, GeneMarkerV2.2.0 software to calculate the relative size of the amplified product fragments and genotyping samples. Results DNA was extracted from all eight bone samples. The average concentration of DNA was 25 ng / μL and D260 / D280 was between 1.7 and 1.9. The detection rate of 9 miniSTR loci in the samples ranged from 78% to 100%, the typing patterns were clearer, and some extra bands appeared in some samples. Conclusion In this paper, nine miniSTR loci typing test method can be used for the detection of DNA typing of bone picking materials.