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将聚丙烯酰胺凝胶中DNAS片段于PCR缓冲液中洗脱,加入PCR反应试剂直接扩增目的DNA作进一步分析,DNA片段回收效率为0.89±0.05,本方法简便、快速。 相似文献
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法医常染色体STR分型 总被引:11,自引:2,他引:9
短串联重复序列 (STR)是广泛存在于人类基因组中的一类具有长度多态性的DNA序列。它由 2~ 6个碱基对构成核心序列 ,呈串联重复排列 ,其长度多态性主要由核心序列拷贝数目的变化而产生。一般认为 ,人类基因组平均每 6~ 10kb就有 1个STR基因座 ,为法医个人识别和亲子鉴定提供了高信息基因座的丰富来源。与限制性片段长度多态性产生的DNA指纹相比 ,用聚合酶链反应技术 (PCR)扩增STR基因座产生的DNA纹印具有更高的灵敏度。STR扩增片段较短 (10 0~ 30 0bp) ,对于常见含部分降解DNA的陈旧斑痕 ,PCR扩增ST… 相似文献
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第三讲 法医DNA分型技术与方法 总被引:1,自引:0,他引:1
法医DNA分型利用分子生物学技术检测、分析人类遗传标记 ,进行个体识别和亲子鉴定。 80年代以来 ,法医DNA分型技术跨越了两大步 ,第一代分型是以DNA分子杂交为核心的限制性片段长度多态性分析技术 ;第二代是以聚合酶链反应 (PCR)为基础衍生出的各种DNA标记分析技术。1 限制性片段长度多态性分析技术早年Jeffreys建立的DNA指纹技术本质是DNA限制性片段长度多态性 (RFLP)分析 ,技术核心是DNA分子杂交。DNA指纹图谱的特异性取决于 2个因素 :一是限制性核酸内切酶特异性 ;二是探针特异性。RFLP分析… 相似文献
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线粒体DNA测序分析及其法医学应用 总被引:2,自引:0,他引:2
本文建立一种以γ-32PATP直接标记PCR扩增引物为测序引物、纯化的PCR扩增产物为模板、Taq酶直接测序的方法、对100倒无血缘关系的中国汉族人线粒体DNA主要高变区D环区的328个碱基(第16091-16418位)进行了核酸序列分析,检出86种基因型,出现率最高者为7%;检出81个可变碱基位点。两个非血缘关系的中国汉族人出现完全相同序列的概率为0.0068。通过对2个四代家系.5个三代家系和10个两代家系分析扯实线粒体DNA是母系遗传。 相似文献
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本文分析了降解DNA作PCR分型易失败的原因,提出了因小片断DNA过多,阻碍引物与膜板的复性及阻断引物延伸的新观点。将降解的DNA电泳,切除小片断DNA,电洗脱回收较大片断DNA,成功地对降解DNA进行了PCR分型。本方法简便、快速、有效。 相似文献
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用PCR-SSP法分析中国辽宁汉族HLA-DRB1基因多态性 总被引:3,自引:1,他引:2
应用PCR-SSP方法对辽宁地区159名无关个体进行HLA-DRB1位点基因分型,检出8组等位基因(扩增片段大小为100bp),基因频率范围在0.02201 ̄0.23899,36种可能基因型中检出33种。经x^2检验符合Hardy-Weinberg平衡定律。本地区汉族群体的期望杂合度为85%,观察杂合度为83%,个人鉴别机率(DP)为0.94,非父排除率(EPP)为66%,本法具有简单,快速,结果 相似文献
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PCR是一种灵敏的DNA检测技术,但是,在对生物学检材进行个人识别时,仍然有一定数量的检材直接PCR扩增失败。为了有效地进行个人识别,本文分析了扩增失败的原因以及成功扩增的对策 相似文献
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法医DNA实验室的DNA污染和防范 总被引:3,自引:2,他引:1
DNA污染是产生DNA鉴定结论错误的重要因素,法医DNA实验室要努力去解决这一问题。DNA污染有自身污染、交叉污染、PCR污染3种。法医DNA实验室要采取实验室分区、严格检验操作步骤、对试剂及消耗材料进行质量控制等方法防止发生DNA污染,采取设置对照样本、核查DNA结果、建立DNA排查数据库等方法监测和发现DNA污染。 相似文献
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Gunjan Sinha 《International Review of Law, Computers & Technology》2001,15(1):73-77
Credit-card-size devices will soon analyze DNA right at a crime scene identifying the perpetrators before they can strike again. 相似文献
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Kavlick MF Lawrence HS Merritt RT Fisher C Isenberg A Robertson JM Budowle B 《Journal of forensic sciences》2011,56(6):1457-1463
Successful mitochondrial DNA (mtDNA) forensic analysis depends on sufficient quantity and quality of mtDNA. A real-time quantitative PCR assay was developed to assess such characteristics in a DNA sample, which utilizes a duplex, synthetic DNA to ensure optimal quality assurance and quality control. The assay's 105-base pair target sequence facilitates amplification of degraded DNA and is minimally homologous to nonhuman mtDNA. The primers and probe hybridize to a region that has relatively few sequence polymorphisms. The assay can also identify the presence of PCR inhibitors and thus indicate the need for sample repurification. The results show that the assay provides information down to 10 copies and provides a dynamic range spanning seven orders of magnitude. Additional experiments demonstrated that as few as 300 mtDNA copies resulted in successful hypervariable region amplification, information that permits sample conservation and optimized downstream PCR testing. The assay described is rapid, reliable, and robust. 相似文献
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Trace DNA is often detected on handled items and worn clothing examined in forensic laboratories. In this study, the potential transfer of trace DNA to bedding by normal contact, when an individual sleeps in a bed, is examined. Volunteers slept one night on a new, lower bed sheet in their own bed and one night in a bed foreign to them. Samples from the sheets were collected and analysed by DNA profiling. The results indicate that the DNA profile of an individual can be obtained from bedding after one night of sleeping in a bed. The DNA profile of the owner of the bed could also be detected in the foreign bed experiments. Since mixed DNA profiles can be obtained from trace DNA on bedding, caution should be exercised when drawing conclusions from DNA profiling results obtained from such samples. This transfer may have important repercussions in sexual assault investigations. 相似文献
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A recent ruling in the Crown Court of Northern Ireland, R v. Hoey, [R v Sean Hoey. 2007, Crown Court of Northern Ireland] has raised questions about the validity of one variant of DNA analysis, often termed LCN. The ruling and subsequent discussion also raises questions about what constitutes validation of a technique.This paper examines what can be achieved in a laboratory based validation study against the Daubert standard and against guidance given in the UK. There is a significant discrepancy between what can be achieved and the Daubert standard but much less of a discrepancy against the UK guidance. Much of the difference relates to differences in word usage, definitional difficulties, and a lack of mutual understanding and communication between the judiciary and forensic scientists. This highlights a gap that needs attention. 相似文献
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《Forensic Science International: Genetics Supplement Series》2019,7(1):229-231
A central question is ‘how did DNA get there’? To help answer this, we visually monitored and recorded DNA transfer from one substrate to another. When an individual touches a substrate, traces of their DNA are transferred (primary/direct) which can then subsequently be transferred to a second substrate (secondary/indirect). Currently DNA transfer and how much remains can only be determined by collecting the biological material from the substrate, isolating the DNA and quantifying the amount recovered. However, Diamond™ Dye (DD) enables such DNA transfer events to be visualised by monitoring the movement of cellular material.We examined primary and secondary DNA transfer using aluminium as a primary substrate with cotton, polyester, aluminium and plastic as secondary substrates and four contact types between two substrates (passive, pressure, friction and friction with pressure). Participants pressed their index finger against the aluminium for 15 s and then DD was applied to the area of contact; cellular material was detected via a fluorescence microscope. Contact between that substrate and a second substrate was performed, using one of the four contact types. After this contact between substrates each was viewed microscopically and transfer of cellular material was recorded.Cellular material could be recorded as having transferred from one substrate to another. Substrate and contact type had an effect on the extent DNA transfers. DNA transferred at a high rate with aluminium as a primary substrate and cotton, polyester and aluminium as secondary substrates when pressure with friction was applied. This information expands our understanding of how DNA transfers and which factors affect it, thus assisting greatly with activity level reporting as to how DNA came to be where it was found. 相似文献