尼日利亚拉各斯吸烟者和非吸烟者MT-loop DNA片段的质量可能用于法医生物学

J. Minari
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引用次数: 0

摘要

线粒体DNA (mtDNA)是一种小的圆形DNA,负责性状的传递。在法医生物学中,mtDNA分析的高灵敏度使法医科学家能够从与犯罪现场相关的证据中获得信息。本研究旨在调查吸烟者和非吸烟者的mtDNA片段,并确定吸烟在多大程度上影响样本人群的mtDNA质量。从酒吧中提取了25个烟头样本,用棉签棒从非吸烟者的唾液中提取了25个样本。采用酶联免疫试剂盒提取线粒体DNA (mtDNA),用分光光度计检测提取的mtDNA的浓度和纯度。使用以下引物序列,在梯度热循环器中进行聚合酶链反应(PCR)以确定mtDNA的高变区。琼脂糖凝胶电泳采用100碱基对阶梯法测定扩增子大小。非吸烟者唾液提取液的DNA纯度(A260/280 2.06 ~ 1.82)高于吸烟者唾液提取液(A260/280 1.82 ~ 1.0)。非吸烟者唾液中的DNA浓度(26.2 ~ 3.0 ng/µl)高于吸烟者唾液中的DNA浓度(26.2 ~ 2.23ng/µl)。琼脂糖凝胶电泳显示扩增的mtDNA高变区大小在295-300碱基对(bp)之间。本研究表明,吸烟者和非吸烟者的mtDNA高变区具有相同的核苷酸碱基对(bp)范围。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
The Quality of MT-loop DNA Segment of Smokers and Nonsmokers in Lagos, Nigeria for Possible use in Forensic Biology
Mitochondrial DNA (mtDNA) is a small circular DNA responsible for transmission of traits. In forensic biology, the high sensitivity of mtDNA analysis allows forensic scientists to obtain information from evidence associated with crime scene. This study was carried out to investigate the mtDNA segment of smokers and non-smokers and to determine to what extent smoking affects the quality of the mtDNA in the sample population. Twenty five cigarette butts were obtained from a bar and twenty five samples were also obtained from saliva of non-smokers using swab stick. Mitochondrial DNA (mtDNA) was extracted from individual samples using zymo kit, spectrophotometer was used to check for the concentration and purity of the extracted mtDNA. Polymerase chain reaction (PCR) was carried out in a gradient thermocycler to ascertain the hypervariable region of the mtDNA using the following primer sequence. Agarose gel electrophoresis was carried out to know the amplicon size using 100 base pair of ladder. The DNA purity on saliva extract for non-smokers was found to be higher (A260/280 2.06 - 1.82) than the purity of saliva from smokers (A260/280 1.82-1.0). The concentration of DNA found on the saliva traces from non-smokers was higher (26.2 - 3.0 ng/µl) than those extracts from smokers (26.2 - 2.23ng/µl). DNA bands obtained from agarose gel electrophoresis showed amplification of the hypervariable region of mtDNA size ranges from 295-300 base pair (bp). This study showed that the hypervariable region of the mtDNA of both smokers and non-smokers have the same range of nucleotide base pair (bp).
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