Investigation of the efficiency of whole genome amplification prior to short tandem repeat analysis using degraded DNA
Investigation of the efficiency of whole genome amplification prior to short tandem repeat analysis using degraded DNA
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使用降解 DNA 进行短串联重复分析之前研究全基因组扩增的效率
DOI：
10.1016/j.fsigss.2019.10.100
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发表时间：
2019
影响因子：
--
通讯作者：
Kazuhiko Kibayashi
中科院分区：
文献类型：
--
作者：
Mitsuyo Machida;Kazuhiko Kibayashi
短串联重复序列（STR）分析容易失败，因为DNA经常被各种环境因素降解。增加起始模板的数量可以提高STR分析的成功率。增加DNA模板数量的一种方法是全基因组扩增（WGA）;然而，很少有研究表明WGA可以用于法医学中降解的DNA样品。因此，我们进行了一个基于PCR的WGA称为“改进的引物延伸预扩增”（mIPEP）之前，降解DNA的STR分析，因为这种方法是受DNA的数量和质量比其他approaches. Salives从四个志愿者干燥滤纸。这些样品用UVA光（365 nm）照射3、7、14和30天。在DNA提取后，使用5、0.5和0.05 ng DNA启动mIPEP方法。在mIPEP之后，使用AmpFlSTR Identifiler PCR扩增试剂盒进行STR分析。该研究得到东京女子医科大学伦理委员会的批准。在没有施加辐射的情况下，在0.05 ng DNA样品中，使用mIPEP制备的样品中可检测到的STR基因座的数量比不使用mIPEP制备的样品中的多。照射3天后，在0.5 ng DNA样品中可检测到的STR基因座的数量在用mIPEP处理的样品中比在没有mIPEP的样品中更少。照射7天和14天后，有和没有mIPEP的可检测STR基因座的数量之间没有显着差异。然而，30天的照射后，在5 ng的DNA样品与mIPEP制备的可检测的STR基因座的数量大于在没有mIPEP。因此，在中度降解的样品中，mIPEP对于STR基因座检测是不必要的，但它确实提高了严重降解样品中STR分析的成功率。
Short tandem repeat (STR) analysis is prone to failure because DNA is frequently degraded by various environmental factors. Increasing the number of starting templates may improve success in STR profiling. One approach to increase the number of DNA templates is whole genome amplification (WGA); however, few studies have demonstrated that WGA can be used on degraded DNA samples in forensics. Therefore, we performed a PCR-based WGA termed “modified improved primer extension preamplification” (mIPEP) prior to STR analysis of degraded DNA, since this method was less affected by DNA quantity and quality than other approaches are.Saliva from four volunteers was dried onto filter papers. These samples were irradiated with UVA light (365 nm) for 3, 7, 14, and 30 days. The mIPEP method was initiated using 5, 0.5, and 0.05 ng of DNA after DNA extraction. Following mIPEP, STR analysis was performed using the AmpFlSTR Identifiler PCR Amplification Kit. This study was approved by the ethics committee at Tokyo Women’s Medical University.The number of detectable STR loci in 0.05 ng DNA sample was greater in those prepared by mIPEP than those prepared without mIPEP, when no radiation was applied. After 3 days of irradiation, the number of detectable STR loci in 0.5 ng DNA sample was smaller in those treated with mIPEP than in samples without mIPEP. There was no significant difference between the number of detectable STR loci with and without mIPEP after 7 and 14 days of irradiation. However, after 30 days of irradiation, the number of detectable STR loci in 5 ng DNA samples prepared with mIPEP was greater than in those without mIPEP. Hence, mIPEP was not necessary for STR loci detection in moderately degraded samples, but it did improve the success of STR profiling in severely degraded samples.
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
用于法医目的的降解和非核DNA的整个基因组扩增。
影响因子：
2.1
作者：
Maciejewska A;Jakubowska J;Pawłowski R
通讯作者：
Pawłowski R
影响因子：
1.5
作者：
Ambers, Angie;Turnbough, Meredith;Budowle, Bruce
通讯作者：
Budowle, Bruce