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通过直接PCR从人毛发根部进行单管基因分型。

Single-tube genotyping from a human hair root by direct PCR.

作者信息

Hayashida Mariko, Iwao-Koizumi Kyoko, Murata Shigenori, Kinoshita Kenji

机构信息

School of Pharmaceutical Sciences, Mukogawa Women's University, Koshien, Nishinomiya 663-8179, Japan.

出版信息

Anal Sci. 2009 Dec;25(12):1487-9. doi: 10.2116/analsci.25.1487.

Abstract

We have developed a simple, labor-saving, inexpensive and rapid SNP genotyping method that directly uses a human hair root as the template. This single-tube genotyping method was used to successfully and reliably genotype the ADH1B and ALDH2 polymorphisms using a hair root (without DNA isolation) and the polymerase chain reaction (PCR) enzyme kit KOD FX. Since the DNA extraction step was eliminated, the possibility of sample contamination was considerably decreased. The single-tube SNP genotyping was performed by coupling the PCR enzyme kit with allele-specific primer (ASP)-PCR. In the post-genome era, a simple and inexpensive method for diagnostic analysis is in high demand, and this PCR method with a hair root as a template will be very useful for genetic diagnoses in biological and medical laboratories.

摘要

我们开发了一种简单、省力、廉价且快速的单核苷酸多态性(SNP)基因分型方法,该方法直接将人类发根用作模板。这种单管基因分型方法利用发根(无需进行DNA分离)和聚合酶链反应(PCR)酶试剂盒KOD FX,成功且可靠地对乙醇脱氢酶1B(ADH1B)和乙醛脱氢酶2(ALDH2)基因多态性进行了基因分型。由于省去了DNA提取步骤,样本污染的可能性大幅降低。单管SNP基因分型通过将PCR酶试剂盒与等位基因特异性引物(ASP)-PCR相结合来进行。在后基因组时代,对简单且廉价的诊断分析方法需求很高,这种以发根为模板的PCR方法在生物和医学实验室的基因诊断中将非常有用。

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