Whole genome amplification of degraded and nondegraded DNA for forensic purposes.
Whole genome amplification of degraded and nondegraded DNA for forensic purposes.
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用于法医目的的降解和非核DNA的整个基因组扩增。
DOI：
10.1007/s00414-012-0764-9
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发表时间：
2013-03
影响因子：
2.1
通讯作者：
Pawłowski R
中科院分区：
文献类型：
--
作者：
Maciejewska A;Jakubowska J;Pawłowski R
降解的DNA经常在法医遗传学实验室中进行分析。对降解DNA进行可靠分析非常重要，因为其结果会影响专家证词的质量和可靠性。最近，许多全基因组扩增（WGA）方法已被提议作为预扩增工具。它们的工作前提是能够从少至单个细胞的DNA量产生微克数量的DNA。我们选择、研究并比较了七种WGA方法，以评估它们“恢复”降解和未降解DNA的能力：简并寡核苷酸引物PCR、引物延伸预扩增PCR、GenomePlex™ WGA商业试剂盒（西格玛）、多重置换扩增、GenomiPhi™扩增试剂盒（阿莫仙生物科学）、限制和环化辅助滚环扩增以及平端连接介导的WGA。使用SGMPlus、YFiler、线粒体DNA和Y染色体单核苷酸多态性分型对这些方法的效率和可靠性进行了分析和比较。对于未降解的DNA，GenomiPhi和PEP方法获得了最佳结果。在降解DNA（200bp）的情况下，GenomePlex获得了最佳结果，它还成功扩增了严重降解的DNA（100bp），从而能够正确分型线粒体DNA和Y - SNP位点。WGA在法医遗传学中对低拷贝数DNA或降解DNA的分析可能非常有用，特别是在引入一些改进措施（样本混合和重复DNA分型）之后。
本文的在线版本（doi:10.1007/s00414 - 2012 - 0764 - 9）包含补充材料，授权用户可获取。
Degraded DNA is often analyzed in forensic genetics laboratories. Reliable analysis of degraded DNA is of great importance, since its results impact the quality and reliability of expert testimonies. Recently, a number of whole genome amplification (WGA) methods have been proposed as preamplification tools. They work on the premise of being able to generate microgram quantities of DNA from as little as the quantity of DNA from a single cell. We chose, investigated, and compared seven WGA methods to evaluate their ability to “recover” degraded and nondegraded DNA: degenerate oligonucleotide-primed PCR, primer extension preamplification PCR, GenomePlex™ WGA commercial kit (Sigma), multiple displacement amplification, GenomiPhi™ Amplification kit (Amersham Biosciences), restriction and circularization-aided rolling circle amplification, and blunt-end ligation-mediated WGA. The efficiency and reliability of those methods were analyzed and compared using SGMPlus, YFiler, mtDNA, and Y-chromosome SNP typing. The best results for nondegraded DNA were obtained with GenomiPhi and PEP methods. In the case of degraded DNA (200 bp), the best results were obtained with GenomePlex which successfully amplified also severely degraded DNA (100 bp), thus enabling correct typing of mtDNA and Y-SNP loci. WGA may be very useful in analysis of low copy number DNA or degraded DNA in forensic genetics, especially after introduction of some improvements (sample pooling and replicate DNA typing). The online version of this article (doi:10.1007/s00414-012-0764-9) contains supplementary material, which is available to authorized users.
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影响因子：
2.2
作者：
Gill, P;Whitaker, J;Buckleton, J
通讯作者：
Buckleton, J
影响因子：
1.6
作者：
Barber, Amy L.;Foran, David R.
通讯作者：
Foran, David R.
影响因子：
2.9
作者：
Hanson, EK;Ballantyne, J
通讯作者：
Ballantyne, J
DOI：
10.1073/pnas.89.13.5847
发表时间：
1992-07-01
影响因子：
11.1
作者：
ZHANG, L;CUI, XF;ARNHEIM, N
通讯作者：
ARNHEIM, N
影响因子：
2.2
作者：
Bender, K;Schneider, PM;Rittner, C
通讯作者：
Rittner, C