Rucinski Cynthia, Malaver Ayda L, Yunis Emilio J, Yunis Juan J
Grupo de identificación Humana e Inmunogenética, Universidad Nacional de Colombia, Bogotá, D.C., Colombia.
J Forensic Sci. 2012 May;57(3):706-12. doi: 10.1111/j.1556-4029.2011.02012.x. Epub 2011 Dec 28.
The quality and efficiency of a standard organic DNA isolation method and a silica-based method using the QIAGEN Blood Maxi Kit were compared to obtain human DNA and short tandem repeats (STRs) profiles from 39 exhumed bone samples for paternity testing. DNA samples were quantified by real-time PCR, and STR profiles were obtained using the AmpFlSTR(®) Identifiler(®) PCR amplification kit. Overall, the silica-based method recovered less DNA ranging from 0 to 147.7 ng/g (average 7.57 ng/g, median = 1.3 ng/g) than did the organic method ranging from 0 to 605 ng/g (average 44.27 ng/g, median = 5.8 ng/g). Complete profiles (16/16 loci tested) were obtained from 37/39 samples (95%) using the organic method and from 9/39 samples (23%) with the silica-based method. Compared with a standard organic DNA isolation method, our results indicate that the published silica-based method does not improve neither the quality nor the quantity of DNA for STR profiling.
为了从39份用于亲子鉴定的挖掘出的骨样本中获取人类DNA和短串联重复序列(STRs)图谱,对一种标准有机DNA提取方法和使用QIAGEN Blood Maxi试剂盒的基于硅胶的方法的质量和效率进行了比较。通过实时PCR对DNA样本进行定量,并使用AmpFlSTR(®) Identifiler(®) PCR扩增试剂盒获得STR图谱。总体而言,与有机方法相比,基于硅胶的方法回收的DNA较少,范围为0至147.7 ng/g(平均7.57 ng/g,中位数 = 1.3 ng/g),有机方法回收的DNA范围为0至605 ng/g(平均44.27 ng/g,中位数 = 5.8 ng/g)。使用有机方法从37/39个样本(95%)中获得了完整图谱(检测16/16个位点),而基于硅胶的方法从9/39个样本(23%)中获得了完整图谱。与标准有机DNA提取方法相比,我们的结果表明,已发表的基于硅胶的方法在用于STR分析时,既没有提高DNA的质量也没有提高其数量。