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使用电泳和凝胶固定化丙烯酰胺探针进行突变分型。

Mutation typing using electrophoresis and gel-immobilized Acrydite probes.

作者信息

Kenney M, Ray S, Boles T C

机构信息

Mosaic Technologies, Inc., Boston, MA, USA.

出版信息

Biotechniques. 1998 Sep;25(3):516-21. doi: 10.2144/98253pf01.

Abstract

A new electrophoresis technology for hybridization-based sequence detection and mutation typing is described. Intrinsic to this approach is copolymerization of specially modified oligonucleotide probes directly into polyacrylamide gels. Electrophoresis of single-stranded samples through gels containing specific immobilized probes results in hybridization-mediated capture of complementary targets. By increasing gel temperature or including denaturants in the buffer, the method can be used to type single-nucleotide polymorphisms. The method can easily be adapted to type mutations in PCR-amplified samples. Acrydite gel technology will also be useful for many other applications, including hybridization-based diagnostics, analysis of gene expression and purification of nucleic acids from biological samples.

摘要

本文描述了一种用于基于杂交的序列检测和突变分型的新型电泳技术。该方法的核心是将经过特殊修饰的寡核苷酸探针直接共聚到聚丙烯酰胺凝胶中。通过含有特定固定探针的凝胶对单链样品进行电泳,可实现杂交介导的互补靶标的捕获。通过提高凝胶温度或在缓冲液中加入变性剂,该方法可用于单核苷酸多态性的分型。该方法可轻松适用于对PCR扩增样品中的突变进行分型。丙烯酰胺凝胶技术还将在许多其他应用中发挥作用,包括基于杂交的诊断、基因表达分析以及从生物样品中纯化核酸。

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