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Chelex法和两种磁珠法提取接触DNA效果的比较 总被引:1,自引:0,他引:1
目的比较Chelex法、DNA IQ磁珠法、EQ国产磁珠法对接触DNA的提取效果。方法将稀释为10ng、100ng的标准品DNA,分别采用Chelex法、DNA IQ磁珠法、EQ国产磁珠法处理;对30例烟蒂和30例牙刷分别采用Chelex法、DNA IQ磁珠法和EQ国产磁珠法提取DNA,然后进行PCR定量和STR检测。结果Chelex法对DNA的提取无损失,DNA IQ磁珠法、EQ国产磁珠法对DNA的提取均有不同程度的损失;烟蒂、牙刷等检材采用Chelex法提取的接触DNA量和IPC CT值显著高于IQ磁珠法、EQ国产磁珠法,但STR检验成功率却低于IQ磁珠法、EQ国产磁珠法。2种磁珠法提取的DNA量、IPC CT值和STR检验成功率无显著性差异。结论污染轻、杂质少的接触DNA检材,用Chelex法提取最为方便快捷;IQ磁珠法、EQ国产磁珠法更适合污染接触DNA检材的提取及自动化操作。 相似文献
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DNA IQ磁珠法结合Maxwell~(TM) 16自动仪提取接触DNA 总被引:1,自引:0,他引:1
目的研究DNA IQ磁珠法结合MaxwellTM 16自动仪对接触DNA提取的应用价值。方法 151份案件接触DNA检材95℃裂解后,采用DNA IQ磁珠法结合MaxwellTM 16自动仪提取DNA,然后进行DNA定量和STR分型检测,统计各种类型的接触DNA含量I、PC CT值和STR分型成功率。结果 151份案件接触DNA检材中,除果核平均DNA获得量为9.51ng以外,其它接触检材的平均DNA获得量均大于10ng,烟蒂检验成功率最高为93%,果核检验成功率较低,为60%。所有DNA样品的IPC CT值均在27左右,纯度高。结论大部分接触DNA检材采用DNA IQ磁珠法结合MaxwellTM 16自动仪可提取到足以进行STR分型的DNA。 相似文献
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在日常检案中,DNA检验人员通常根据尸体的腐败程度来决定提取那种检材以及采取何种方法得到足够的遗传信息。据高俊薇等[1]报道,当检验腐败尸体的心血、肌肉等组织无法得到STR分型结果时,拔取其指甲进行STR分型往往能得到满意的效果。指甲检验成功率与保存环境有关,土埋1个月以上尸体的指甲DNA经有机法提取成功率不高。本例在处理一起土埋2年半、完全白骨化的尸体时,以指甲作为检验对象,分别以有机法、硅珠法和磁珠法对经Chelex-100提取后的模板DNA进行纯化浓缩,并对其效果进行比较。一、样本来源2005年7月中旬,在距本市某内陆河入海… 相似文献
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在日常检案中,DNA检验人员通常根据尸体的腐败程度来决定提取那种检材以及采取何种方法得到足够的遗传信息。据高俊薇等报道。当检验腐败尸体的心血、肌肉等组织无法得到STR分型结果时,拔取其指甲进行STR分型往往能得到满意的效果。指甲检验成功率与保存环境有关.土埋1个月以上尸体的指甲DNA经有机法提取成功率不高。本例在处理一起土埋2年半、完全白骨化的尸体时,以指甲作为检验对象,分别以有机法、硅珠法和磁珠法对经Chelex-100提取后的模板DNA进行纯化浓缩。并对其效果进行比较。 相似文献
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目的 用mtDNA中细胞色素氧化酶辅酶Ⅱ(COⅡ)基因序列鉴定法医学中常见食尸性苍蝇及其幼虫的种类. 方法 收集郑州地区大白鼠尸体上的苍蝇及其幼虫,提取DNA,PCR扩增CO Ⅱ序列,测序,用Clustalx和MEGA 4.0软件对基因序列进行比对分析及构建系统进化树.结果 成虫与幼虫基因差异不明显,CO Ⅱ基因序列可以对棕尾别麻蝇、巨尾阿丽蝇和亮绿蝇进行鉴定,铜绿蝇与丝光绿蝇进化距离较近,CO Ⅱ序列不能将他们区分开,同时还发现巨尾阿丽蝇和亮绿蝇存在种群单核苷酸多态性. 结论 mtDNA中COⅡ序列能有效鉴定郑州地区部分常见食尸性苍蝇的种类,方法 简便、准确. 相似文献
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《Forensic Science International: Genetics Supplement Series》2013,4(1):e121-e122
The analysis of a biological samples found in crime scenes can be a challenging task. Minute amount of DNA, degraded DNA and the presence of inhibitors in biological samples, can interfere with the achievement of a complete genetic profile.Chelating resin, silica membranes and silica-coated with paramagnetic resin were the techniques used in this study for the isolation and purification of DNA. Our aim was to find out the best DNA extraction method for application in the presence of biological samples buried in clay soil. 相似文献
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低拷贝模板DNA分析技术研究进展 总被引:1,自引:0,他引:1
近年来,低拷贝(LCN)模板类生物物证在法医DNA分析中占有了越来越重要的地位。用于低拷贝模板DNA的检测方法也得到飞速发展。本文通过对各种LCN-DNA分析技术如增加扩增循环数、纯化扩增产物、全基因组扩增、激光捕获显微切割等检测方法的综述,以及对LCN—DNA检测结果的分析评价,全面介绍LCN分析技术在法庭科学应用的最新进展及存在问题。最大限度地拓展低拷贝模板类生物物证在刑事司法领域的应用。 相似文献
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《Forensic Science International: Genetics Supplement Series》2019,7(1):120-122
Current sampling strategy for laboratories typing bones for human identification include samples obtained from femur, tooth and temporal bone. Latest studies suggest that the small bones of the hands and feet were very similar or even better in DNA yield. These bones can be easily sampled with a disposable scalpel and thus reduce potential DNA contamination. The aim of our study was to determine the suitability of metatarsals, metacarpals and phalanges for genetic identification. 48 bone samples from 8 different skeletons (six from 18th century and two from 3rd century) were obtained from 5 archaeological sites in Slovenia. In each skeleton, 6 different skeletal elements were sampled (temporal bone, molar, femur, metacarpal bone, metatarsal bone and proximal phalanx of the hand), and strict precautions followed to prevent contamination. Half of gram of bone powder was decalcified using full demineralization extraction method. The DNA was purified in a Biorobot EZ1 (Qiagen), DNA content determined with the PowerQuant kit (Promega), and autosomal STR typing performed with the Investigator ESSplex Plus kit (Qiagen). Up to 8.75 ng DNA/g of powder was obtained from samples analyzed. The highest yields were detected in temporal bone and the lowest in femur. The success rate of STR typing was evaluated according to the number of successfully typed loci and a strong correlation between the success rate of STR typing and the amount of extracted DNA was confirmed. For all eight skeletons full consensus genetic profiles were determined from skeletal elements analyzed. Our findings suggest it would be suitable to include metatarsal and metacarpal bones in sampling strategy for human identification although further research is needed to substantiate the findings of this study. 相似文献
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目的分析死后不同时间大鼠视网膜细胞核DNA含量变化图像,探讨视网膜细胞核DNA降解与死亡时间(PMI)的关系。方法90只成年健康雌性SD大鼠,随机分成15组,死后0—28h内(20℃),每2h提取视网膜细胞进行Feulgen-Vans染色;采用图像分析系统检测视网膜细胞核的异形指数(ID)、积分光密度(IOD)和平均光密度(AOD),并运用SPSS12.0软件对测量数据进行线性回归分析。结果视网膜细胞核AOD和IOD随死亡时间的延长逐渐下降,ID则呈上升趋势。28h内各参数变化的回归方程如下:YAOD=-0.009XAOD+0.590,R^2=0.949,Y100=-0.097X。+18.903,R^2=0.968,Y10=0.122X10+2.246,R^2=0.951。结论大鼠死后视网膜细胞核DNA含量随着死亡时间的延长而呈规律性降解,并与PMI具有良好的相关性。 相似文献
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Li Yen Candy Lee PhD Yong Sheng Lee BSc Jiayu Tan BSc Jun Yu Lee BSc Christopher Kiu-Choong Syn PhD 《Journal of forensic sciences》2024,69(2):430-436
The ability to detect low level DNA brings with it the uncertainty of whether the detected DNA is a result of transfer. To address this uncertainty, a simulation study was conducted in which a mock illicit drug packet was placed into the personal bags of individuals. When the average transit time of the packets was increased from around 2 h to more than 14 h, the percentage of the DNA profiles recovered from the packets which could be attributed to the individuals increased greatly from 5.3% to 48.6%. We found that drug packers who were poor shedders could not be included as contributors to the DNA profiles from the drug packets at all and there was a higher chance that individuals other than themselves could be included as contributors to the DNA profile recovered from drug packets. We also found that it was equally likely that the drug packers who had direct contact with the drug packets and bag owners who did not, could be included as contributors to the DNA profiles recovered from the packets. The results in this study highlight the importance of taking into consideration the transit time of drug packet, the shedder status of the alleged packer and the history of an item, when evaluating DNA evidence in the context of illicit drug activities. 相似文献
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图像分析技术测定DNA含量变化推断死亡时间的研究及应用前景 总被引:7,自引:1,他引:6
图像分析技术是20世纪50年代发展起来的一门技术,它通过对图像信息的收集、处理、计算、分析而得出图像各部分的数量变化,在生物医学领域已经成为一种常规的定量技术。本文对图像分析技术测定DNA含量的原理、死后细胞核DNA降解的规律及图像分析技术定量DNA推断死亡时间的应用前景进行了综述。 相似文献
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O. Santapa S. Filippini S. Valente M.B. Rodriguez Cardozo 《Forensic Science International: Genetics Supplement Series》2009,2(1):147-148
Analysis of forensic samples to evaluate the rate of success for molecular markers: autosomal STRs, Y chromosome, and mitochondrial DNA. Since 2006 to date a total of 390 forensic samples were analyzed: bones, teeth, hairs, swabs, stains and paraffin embedded tissue. Bones and teeth, were pulverized in a Freezer Mill, extracted by chloroform/phenol/isoamyl alcohol method, and then purified with Centricon 100 columns. DNA from paraffin was extracted with QIAmp DNA Mini kit (QIAGEN). Mitochondrial DNA Control Region sequences were determined for regions HV1/HV2. Sequencing was performed using the BigDye® Terminator v 1.1 Kit and analyzed in ABIPRISM® 3100 Genetic Analyzer (AB). STRs were amplified using Amp FlSTR Identifiler®, Minifiler® and YFiler® Kit (AB) and analyzed in ABI PRISM® 3100 Genetic Analyzer and ABI PRISM® 3130xl Genetic Analyzer (AB). Among forensic samples, bones and teeth analyzed for autosomal STRs, we obtained successful results in all of them. Incomplete typing are represented by loci of higher molecular weight, which demonstrates the poor quality of the sample due to its state of degradation and obtained better results using mini STRs. Successful results in sequencing for mitochondrial HV1 region for all samples analyzed, but in few hair samples we obtained mixed sequences and that represented important difficulties for the analysis. Age of samples and conservation are factors related which affect DNA viability. Autosomal STRs solved all the samples analyzed in our study, but Y chromosome analysis and mitochondrial DNA sequencing are also important and necessary markers in some forensic cases. 相似文献