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作 者:李成志[1] 任甫[1] 席焕久[1] 李文慧[1] 朱博[1] 徐国昌[2]
机构地区:[1]辽宁医学院生物人类学研究所法医教研室,辽宁锦州121001 [2]南阳理工学院生物人类学研究所,河南南阳473004
出 处:《中国法医学杂志》2014年第5期409-412,416,共5页Chinese Journal of Forensic Medicine
基 金:辽宁省百千万人才工程资助项目(2010921042);河南省科技厅基础与前沿技术研究类项目(132300410088)
摘 要: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 type the 9 miniSTR loci(D20S1082, D6S474, D12ATA63, D9Sl122, D2S1776, D1S1627, D3S4529, D2S441, Amelogenin) in femoral bones incinerated at 300℃ respectively. Methods The 8 samples of femoral bones were incinerated at 300℃. DNA was extracted from burned bones by improved phenol-chloroform. Amplification of 9 miniSTR loci was performed on the gradient PCR respectively. Data were collected by the 3130 gene analyzer, the length of the amplification fragments was calculated and the genotyping was made using GeneMarker V2.2. 0 software. Results In the femoral bones incinerated at 300℃, DNA can be extracted successfully in each sample and the average concentration of DNA was 25ng/μL,and its value of D260/D250 was between 1.7 and 1.9. The detection rate of 9 miniSTR loci was between 78% and 100% in the 8 samples. All the typing graphs were clear, but sometimes an additional profile appeared in one sample. Conclusion The way of detection and genotyping for this 9 miniSTR loci can be used for the DNA analysis of burned bones.
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