基于TaqMan法的人DNA定量试剂盒的比较

Takashi Fukagawa, Haruhiko Watahiki, Yusuke Mita, Tetsushi Kitayama, K. Fujii, N. Mizuno
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引用次数: 0

摘要

人类DNA定量是后续短串联重复(STR)分析和解释的重要步骤,因为它可以提供提取DNA的数量和质量的各种信息。在这项研究中,我们比较了四种市售的基于TaqMan检测的定量阳离子试剂盒和一种基于插层染料的定量阳离子试剂盒。我们研究了以下三点:1)使用GlobalFiler和Y - ler Plus PCR扩增试剂盒扩增1 ng DNA时个体间平均峰高的变化,每个定量检测试剂盒测定的数量为1 ng; 2)雄性DNA定量检测中女性DNA的存在eŠect; 3)“降解指数”与STR电泳之间的关系。使用GlobalFiler产生的平均峰高在四种基于TaqMan的试剂盒中显示出比基于插层染料的试剂盒更少的个体依赖性变化。使用Y + ler Plus生成的平均峰高在能够定量男性DNA和总人类DNA的三种TaqMan试剂盒中显示出相似的变化。在三个试剂盒中,大量雌性DNA的存在对雄性DNA的定量阳离子几乎没有eŠect影响。当对高度降解的DNA样本进行定量分析时,三个试剂盒的“降解指数”diŠered显著不同。这可能是由于试剂盒中长片段目标的diŠerent扩增子大小。在案例工作中实施新的人类DNA定量试剂盒之前,有必要了解所采用的定量试剂盒的特性。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Comparison of human DNA quantification kits based on a TaqMan assay
Human DNA quantiˆcation is an important step for subsequent short tandem repeat (STR) analysis and interpretation because it can provide various information on the quantity and quality for the extracted DNA. In this study, we compared four commercially available quantiˆcation kits based on a TaqMan assay and an intercalating-dye based quantiˆcation kit. The following three points were investigated: 1) variations of the average peak heights among individuals when 1 ng of DNA, the amount being determined by each quantiˆcation kit, was ampliˆed using GlobalFiler and Yˆler Plus PCR Ampliˆcation Kits, 2) eŠect of the presence of a female DNA on the quantiˆcation of male DNA, and 3) relationship between ``Degradation Index'' and the STR electropherograms. The average peak heights generated using GlobalFiler showed less individual-dependent variations in the four TaqMan based kits than the intercalating-dye based kit. The average peak heights generated using Yˆler Plus showed similar variations among the three TaqMan based kits capable of male DNA quantiˆcation along with total human DNA. The presence of a large amount of female DNA had little eŠect on male DNA quantiˆcation in all the three kits. When highly degraded DNA samples were quantiˆed, the ``Degradation Indices'' diŠered signiˆcantly among three kits. It was probably due to the diŠerent amplicon sizes of the long fragment targets among kits. Before implementing a new human DNA quantiˆcation kit in casework, it is essential to understand the characteristics of the adopted quantiˆcation kit.
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