通过毛细管电泳分析聚合酶链反应产物及其在基因单碱基取代检测中的应用。
Analysis of polymerase chain reaction product by capillary electrophoresis and its application to the detection of single base substitution in genes.
作者信息
Arakawa H, Uetanaka K, Maeda M, Tsuji A
机构信息
School of Pharmaceutical Sciences, Showa University, Tokyo, Japan.
出版信息
J Chromatogr A. 1994 Mar 25;664(1):89-98. doi: 10.1016/0021-9673(94)80632-2.
Capillary gel electrophoresis (CGE) was studied for the direct analysis of polymerase chain reaction (PCR) amplified samples. A low cross-linked polyacrylamide gel (3%T, 0.5%C) was used for CGE with treated and untreated silica capillaries. CGE showed high reproducibility and resolution in the separation of DNA fragments (ca. 100-1000 base pairs) produced by PCR. The CGE system was applied to the detection of an amplification refractory mutation system (ARMS) and PCR-restriction fragment length polymorphism (PCR-RFLP), which are detection methods of single base substitution in genes using PCR. With the CGE system, full automation of PCR product detection is feasible.
研究了毛细管凝胶电泳（CGE）用于直接分析聚合酶链反应（PCR）扩增样品。采用低交联聚丙烯酰胺凝胶（3%T，0.5%C），在经过处理和未处理的硅胶毛细管上进行CGE。CGE在分离由PCR产生的DNA片段（约100 - 1000个碱基对）时显示出高重现性和分辨率。该CGE系统应用于扩增不应性突变系统（ARMS）和PCR - 限制性片段长度多态性（PCR - RFLP）的检测，这两种方法是利用PCR检测基因中单碱基取代的方法。使用CGE系统，PCR产物检测的完全自动化是可行的。
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