法医学杂志

法医学杂志 ›› 2002, Vol. 0 ›› Issue (3): 155-159.

• 论文 • 上一篇    下一篇

PCR扩增体系的体积减少对DNA分析的影响

周怀谷,平原,徐庆文,胡伟   

  1. 上海市刑事科学技术研究所,上海市刑事科学技术研究所,上海市刑事科学技术研究所,上海市刑事科学技术研究所 上海200083 ,上海200083 ,上海200083 ,上海200083
  • 发布日期:2002-06-25 出版日期:2002-06-28

Effect of PCR reaction volume on the accuracy of human identification tests

ZHOU HUAI-GU,PING YUAN,XU QING-WEN,ET AL.(SHANGHAI INSTITUTE OF CRIMINAL SCIENCE AND TECHNOLOGY,SHANGHAI200083)   

  • Online:2002-06-25 Published:2002-06-28

摘要： 目的探讨PCR扩增体系的体积减少对DNA分析的影响。方法4份样品采用ProfilerPlus试剂盒,在同样条件下,对50μl、25μl、12.5μl、6.25μl四种体积的体系进行扩增,扩增产物分别经ABIPRISMTM310型基因分析仪、ABIPRISMTM377型测序仪、ABIPRISMTM3100型基因分析仪电泳,并经GeneScan3.1基因扫描软件和Genotyper2.5分析软件分析得到实验结果。结果小体积的扩增体系容易出现等位基因的丢失、额外等位基因的产生等现象。结论在检材情况较差时,应该慎用小体积的扩增体系。

关键词: PCR扩增体系, DNA分析, 等位基因丢失, 额外等位基因

Abstract: Objective To investigate the effect of different PCR amplification volume on the accuracy of human identification test.Methods Human genome DNA samples were amplified using ABI PRISM Profiler Plus kits in50 μ l,25 μ l,12.5 μ l,and6.25 μ l reaction volume,respectively.The thermocycle parameters were the same.All PCR products were then electrophoresized on ABI PRISM310Genetic Analyzer,377DNA Sequencer,and3100Genetic Analyzer.Data were processed by ABI PRISM GeneScan and Genotyper software.Results The less re action volume,the more alleles losing or alleles adding observed.Conclusion Non-standard volume of PCR am plification reaction should be used carefully in human identification test,especially when the sample DNA quali ty is not so satisfied.

Key words: PCR amplification system, DNA analysis, allele loss, additional allele