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直接 PCR 与提取法从不同基质中提取的样本生成 DNA 图谱的比较。

A comparison between direct PCR and extraction to generate DNA profiles from samples retrieved from various substrates.

机构信息

Centre for Forensic Science, University of Strathclyde, Department of Pure and Applied Chemistry, Royal College Building, 204 George Street, G1 1XW Glasgow, UK.

出版信息

Forensic Sci Int Genet. 2012 May;6(3):407-12. doi: 10.1016/j.fsigen.2011.08.007. Epub 2011 Sep 16.

Abstract

Direct PCR generates DNA profiles from samples without using the extraction process. During sample extraction, DNA may be lost due to the methods used, which can affect the quality of the DNA profile obtained. This is not the case with direct PCR, where the sample is transferred directly into the PCR tube. Here, we report on the ability of direct PCR to generate DNA profiles from low amounts of control DNA retrieved from various surfaces using PowerPlex 16 HS. A comparison is made with samples undergoing a preliminary extraction stage using QiaAmp DNA Micro kits. Samples subjected to direct PCR generated DNA profiles with higher peak heights and lower allele dropout on all the different substrates tested when compared to the samples subjected to extraction. The amount of DNA retrieved from each substrate also varied even though the same amount of starting material was deposited, proving that the type of substrate can affect the retrieval of DNA.

摘要

直接 PCR 无需提取过程即可从样本中生成 DNA 图谱。在样本提取过程中,由于使用的方法,DNA 可能会丢失,这会影响获得的 DNA 图谱的质量。直接 PCR 则不会出现这种情况,因为样本直接转移到 PCR 管中。在这里,我们报告了直接 PCR 从使用 PowerPlex 16 HS 从各种表面上回收的低量对照 DNA 生成 DNA 图谱的能力。并将其与使用 QiaAmp DNA Micro 试剂盒进行初步提取阶段的样品进行了比较。与经过提取的样品相比,直接 PCR 处理的样品在所有测试的不同基质上生成的 DNA 图谱具有更高的峰高和更低的等位基因缺失。即使沉积了相同量的起始材料,从每个基质中回收的 DNA 量也有所不同,这证明了基质的类型会影响 DNA 的回收。

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