Chelex-100提取生物检材DNA实时PCR定量研究

发布时间：2018-04-15 01:27

本文选题：法医物证学 + 实时定量PCR　；参考：《中国法医学杂志》2008年01期

【摘要】：目的研究Chelex-100法提取的生物检材DNA用量与复合STR分型成功率的关系。方法113份各种生物检材采用Chelex-100法提取DNA,应用Quantifiler人类DNA定量试剂盒在ABI 7500荧光定量PCR仪上进行实时PCR定量,同时用Identifiler复合扩增系统在ABI 3100遗传分析仪上对这些DNA样品进行STR分型。结果各种生物检材提取的DNA浓度分别为:37份滤纸、纱布血痕0.042~5.28ng/μl,16份口腔拭子1.15~4.21ng/μl,18份烟头0.016~1.46ng/μl,10份肋软骨0.531~14.40ng/μl,8份肌肉5.75~24.80ng/μl,7份指甲0.788~11.50ng/μl,17份精斑0.79~99.50ng/μl。在建立的8μl扩增体系中,根据上述结果,调整用于复合STR扩增的DNA模板量在0.5~3ng之间,大部分样品可获得完全的STR分型。结论Chelex-100法提取的检材DNA模板用量在0.5~3ng之间可得到有效STR扩增,浓度为0.5ng/μl以上的DNA样品,用小体积模板(1μl)比大体积(3μl)模板扩增效果好。
[Abstract]:Objective to study the relationship between the amount of DNA extracted by Chelex-100 and the success rate of compound STR typing.Methods Chelex-100 method was used to extract the DNA from 113 biological samples. Quantifiler human DNA quantitative kit was used for real-time PCR quantification on ABI 7500 fluorescence quantitative PCR instrument. STR typing of DNA samples was carried out by Identifiler multiplex amplification system on ABI 3100 genetic analyzer.Results the concentrations of DNA extracted from various biological samples were: 37 pieces of filter paper, 16 samples of 0.042~5.28ng/ 渭 l, 1.15~4.21ng/ 渭 l, 18 0.016~1.46ng/ 渭 l, 10 0.531~14.40ng/ 渭 l, 8 5.75~24.80ng/ 渭 l, 7 0.788~11.50ng/ 渭 l, 17 0.79~99.50ng/ 渭 l.In the established 8 渭 l amplification system, according to the above results, adjusting the amount of DNA template used for compound STR amplification between 0.5~3ng, most samples can obtain complete STR typing.Conclusion the effective STR amplification can be obtained when the amount of DNA template extracted by Chelex-100 method is between 0.5~3ng. The DNA sample with the concentration of more than 0.5ng/ 渭 l can be amplified with 1 渭 l of small volume template than with 3 渭 l of large volume template.
【作者单位】：广州市刑事科学技术研究所广州市刑事科学技术研究所广州市刑事科学技术研究所南方医科大学广东广州广州市刑事科学技术研究所广州市刑事科学技术研究所广州市刑事科学技术研究所
【分类号】：D919

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