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单根毛发的DNA分型

DNA typing from single hairs.

作者信息

Higuchi R, von Beroldingen C H, Sensabaugh G F, Erlich H A

机构信息

Department of Human Genetics, Cetus Corporation, Emeryville, California 94608.

出版信息

Nature. 1988 Apr 7;332(6164):543-6. doi: 10.1038/332543a0.

Abstract

The characterization of genetic variation at the DNA level has generated significant advances in gene and disease mapping, and in the forensic identification of individuals. The most common method of DNA analysis, that of restriction fragment length polymorphism (RFLP), requires microgram amounts of relatively undegraded DNA for multi-locus typing, and hundreds of nanograms for single-locus comparisons. Such DNA frequently cannot be obtained from forensic samples such as single hairs and blood stains, or from anthropological, genetic or zoological samples collected in the field. To detect polymorphic DNA sequences from single human hairs, we have used the polymerase chain reaction (PCR), in which specific short regions of a gene can be greatly amplified in vitro from as little as a single molecule of DNA. We have detected genetically variable mitochondrial and nuclear DNA sequences from the root region of shed, as well as freshly-plucked, single hairs; mitochondrial DNA (mtDNA) sequences have been detected in a sample from a single hair shaft. We have used three different means of DNA typing on these samples: the determination of amplified DNA fragment length differences, hybridization with allele-specific oligonucleotide probes, and direct DNA sequencing.

摘要

DNA水平上的遗传变异特征分析在基因和疾病图谱绘制以及个体的法医鉴定方面取得了重大进展。最常用的DNA分析方法,即限制性片段长度多态性(RFLP)分析,多位点分型需要微克量的相对未降解的DNA,单一位点比较则需要数百纳克。这样的DNA常常无法从法医样本(如单根毛发和血迹)中获得,也无法从野外采集的人类学、遗传学或动物学样本中获得。为了从单根人发中检测多态性DNA序列,我们采用了聚合酶链反应(PCR),在该反应中,基因的特定短区域可以在体外从仅一个DNA分子开始大量扩增。我们已经从脱落毛发的根部以及刚拔下的单根毛发中检测到了遗传可变的线粒体和核DNA序列;在单根毛发轴的一个样本中检测到了线粒体DNA(mtDNA)序列。我们在这些样本上使用了三种不同的DNA分型方法:确定扩增DNA片段长度差异、与等位基因特异性寡核苷酸探针杂交以及直接DNA测序。

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