共查询到17条相似文献,搜索用时 93 毫秒
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用chelex-100提取骨骼DNA的研究 总被引:4,自引:3,他引:1
建立了应用chelex-100加蛋白酶K提取骨骼DNA的方法,并对1~30年31例骨骼进行了实验.该方法操作简单、快速、重复性好,灵敏度以及阳性率高,适用于法医学鉴定. 相似文献
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骨骼DNA提取与检测是大规模灾难受害者、失踪人员以及刑事案件法医DNA鉴定过程的重要组成部分。骨骼的部位、类型以及温度、湿度、微生物、酸碱度等因素都会影响骨骼中DNA的“质”和“量”,从而影响检测的准确性。随着科技的发展,短串联重复序列基因座、DNA检测试剂盒以及单核苷酸多态性技术、线粒体DNA检测技术不断进步,提高了骨骼DNA的提取与检测的成功率。因此,了解影响DNA提取与检测的骨骼类型以及环境因素,选取适当的检测方法和手段显得尤为重要,通过重点综述影响骨骼DNA质量的环境因素以及骨骼DNA提取与检测手段的研究进展,以期为法医学领域使用骨骼作为检材时遇到的问题提供指导。 相似文献
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陈旧骨骼DNA提取及性别鉴定 总被引:9,自引:0,他引:9
用本室建立的方法对3~15年的陈旧骨骼进行DNA提取,并用X、Y同源引物扩增AInelogenin基因X、Y特异性片段,所有检材均得出正确结论,表明该方法快速、灵敏、可靠,适用于陈旧骨骼性别鉴定。 相似文献
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目的探讨建立骨骼及牙齿DNA自动化提取的新方法。方法将33份骨骼及15份牙齿样本分别用冷冻研磨和手工处理两种方法研磨成粉,采用AutoMate ExpressTM自动化法医DNA提取系统提取DNA并定量。结果 AutoMate ExpressTM自动化法医DNA提取系统能够在3h左右完成骨骼、牙齿DNA的提取,两种方法处理的骨骼样本所得DNA质量浓度差异无统计学意义。冷冻研磨处理的骨骼和牙齿样本均获得了较好的STR分型结果,且牙齿样本所得DNA质量浓度高于手工提取所得。结论应用AutoMate ExpressTM自动化法医DNA提取系统是自动化提取骨骼、牙齿DNA的一种新方法,可应用于法医实际案件检验。 相似文献
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目的对纳米磁珠法提取纯化骨骼DNA的效果进行比较评价,为方法选择提供应用参考。方法取泥土掩埋、水中浸泡1~10年不等的25根长骨,经水洗、刮净,液氮冷冻研磨器将骨骼研磨成粉末状,分别应用纳米磁珠提取法和King Fisher仪器自动化提取法提取DNA,IdentifilerPlus试剂盒进行扩增,ABI 3100遗传分析仪进行STR分型检测;对两种方法提取的DNA定量和经扩增、分型检测的结果进行比较。结果骨骼样本采用纳米磁珠法提取到的样本DNA(1.237 5ng/μL±0.319 2ng/μL),较之King Fisher法的浓度(0.506 2ng/μL±0.280 5ng/μL)更高,两种方法间差异具有统计学意义(P0.05);而纳米磁珠法的分型成功率亦更高,两种方法间差异具有统计学意义(P0.05)。结论用纳米磁珠提取纯化骨骼DNA,能得到高质量DNA模板,有利于提高分型检验的成功率,可在实际检案中选择使用。 相似文献
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Stijn Desmyter Christine De Greef 《Forensic Science International: Genetics Supplement Series》2008,1(1):24-25
Eight human bone samples, from a forensic case, were extracted in parallel using our standard protocol with and without PTB in the buffer. Both methods were sometimes inadequate for (complete) STR profiling, while the presence of PTB even decreases the DNA yield.The complete decalcification of the bone extraction residues in an EDTA-solution with SDS recovered sufficient amounts of DNA, which resulted in complete STR profiling for all samples. Complete decalcification without SDS yielded even higher amounts of DNA and also complete STR profiling for all samples.Similar results were obtained for the DNA extraction from a human tooth. 相似文献
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目的建立一种简约且易于自动化操作的Whatman FTA卡血样DNA提取方法。方法取直径1.2mm FTA卡血样,分别用FTA-DNA直接提取法及FTA常规提取方法提取DNA,AB-Identifiler^TM试剂盒分别进行10山和25山体系检测比较,并筛选批量grA卡血样DNA自动化提取程序。结果200份grA卡血样分别采用2种方法提取DNA模板,25μl体系PCR-STR检测,分型结果均良好。10μl体系检测,FTA-DNA直接提取法优于FTA常规提取方法,图谱RFU值为1000~2000,采用自动化工作站运行该提取程序较手工操作DNA分型图谱均衡性更佳;采用FTA常规提取方法检测,图谱RFU值100~2000,小片段优先扩增现象明显,且有19%的样本出现丢峰现象,采用自动化工作站运行该程序对其结果改善不明显。经工作站运行FTA-DNA直接提取法自动程序及10山体系检测本2万余份FTA卡采集的血样,均获得16个STR基因座DNA分型结果。结论本文建立的FTA-DNA直接提取法适用于FTA卡血样自动化DNA建库。 相似文献
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目的人骨骼和牙齿DNA提取方法的比较和优化。方法收集18份不同个体的长骨、30颗磨牙和同一个体2根股骨、8颗磨牙。利用TissueLyser-Ⅱ组织破碎仪和PreFiler Express BTA^TM法医DNA提取试剂盒(BTA法),应用Automate Express^TM自动化法医DNA提取系统提取DNA,进行STR分型,与脱钙法进行比较,并进行实验条件优化。结果用TissueLyser-Ⅱ结合BTA法,约2.5h即可完成骨骼和牙齿的DNA提取,分型成功率分别为94.4%和96.7%。与脱钙法比较,两种方法获得DNA质量浓度和检出率比较接近(P〈0.05),但BTA法在操作过程方面更具优势。最佳样本量为100mg,消化时间为2h。结论采用TissueLyser-Ⅱ组织破碎仪结合BTA法对骨骼和牙齿进行DNA提取和分型检验,能满足实际检案的要求,可在法医学实践中选择使用。 相似文献
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《Forensic Science International: Genetics Supplement Series》2013,4(1):e174-e175
Five DNA extraction systems were assessed for their DNA extraction efficiency on samples of fresh pig bone. Four commercially available silica-based extraction kits (ChargeSwitch® gDNA Plant Kit (Life Technologies), DNA IQTM System Kit (Promega), DNeasy® Blood & Tissue Kit (Qiagen) and PrepFiler® BTA Forensic DNA Extraction Kit (Life Technologies)) and a conventional phenol-chloroform method were tested in this study. Extracted DNA samples were quantitated with GoTaq® qPCR Master Mix (Promega) using an Applied Biosystems® 7500 Real-Time PCR System and the extracts were amplified using an in-house multiplex system. The phenol-chloroform extraction produced higher yields of DNA than the silica-based extraction methods. Among the silica-based extractions ChargeSwitch® gDNA Plant Kit recovered the highest amounts of DNA. However, all methods produced DNA that could be amplified and none of the extracts contained any detectable inhibition. 相似文献
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Hwan Young Lee Myung Jin Park Na Young Kim Jeong Eun Sim Woo Ick Yang Kyoung-Jin Shin 《Forensic Science International: Genetics Supplement Series》2010,4(5):275-280
The recovery of DNA data from old skeletal remains is often difficult due to degraded and very low yield of extracted DNA and the presence of PCR inhibitors. Herein, we compared several silica-based DNA extraction methods from artificially degraded DNA, DNA with PCR inhibitors and DNA from old skeletal remains using quantitative real-time PCR. We present a modified large-scale silica-based extraction combined with complete demineralization, that enables maximum DNA recovery and efficient elimination of PCR inhibitors. This is performed with high concentration of EDTA solution for demineralization of bone powder followed by QIAamp® spin columns and buffers from the QIAquick® PCR purification kit. We have successfully used this modified technique to perform STR analysis for 55-year-old skeletal remains. The results of this study will contribute to solve the forensic cases dealing with skeletal remains. 相似文献