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[使用引物延伸预扩增(PEP)方法进行DNA分型——分型可靠性及检测限研究]

[Typing of DNA using the primer extension preamplification (PEP) method--studies of reliability of typing and detection limits].

作者信息

Satoh K, Takahashi K, Itoh Y, Kobayashi R

机构信息

Criminal Investigation Laboratory, Metropolitan Police Department of Tokyo, Japan.

出版信息

Nihon Hoigaku Zasshi. 1998 Jun;52(3):184-90.

PMID:9780662
Abstract

Primer extension preamplification (PEP), which can amplify sequence-independently a limited quantity of DNA as a whole, allows multiple DNA analyses by polymerase chain reaction (PCR). This technique may be applicable to forensic cases, especially in cases where only small amounts of DNA are available. To define the accuracy and sensitivity of PEP, the DNA typing results of nine loci (TH01, MCT118, HLA-DQ alpha, amelogenin, LDLR, GYPA, HBGG, D7S8, GC) by PCR with PEP (PEP-PCR) were compared with those by PCR without PEP. Both results were identical to each other for each sample tested. Furthermore, amplification of an initial genomic DNA by PEP was found to range from 15 to 600 times of the initial quantity and the efficiency of PEP may depend on the sequences flanking the loci tested. These results indicate that PEP can increase typing potential of PCR from forensic samples.

摘要

引物延伸预扩增(PEP)能够对少量DNA进行全序列非依赖性扩增,从而可通过聚合酶链反应(PCR)进行多种DNA分析。该技术可能适用于法医案件,尤其是在仅获得少量DNA的情况下。为了确定PEP的准确性和灵敏度,将采用PEP的PCR(PEP-PCR)对9个基因座(TH01、MCT118、HLA-DQα、牙釉蛋白、低密度脂蛋白受体、血型糖蛋白A、β-球蛋白、D7S8、Gc)的DNA分型结果与未采用PEP的PCR结果进行了比较。对每个测试样本而言,两种结果均彼此相同。此外,发现通过PEP对初始基因组DNA的扩增范围为初始量的15至600倍,且PEP的效率可能取决于所测试基因座侧翼的序列。这些结果表明,PEP可以提高法医样本PCR的分型潜力。

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