A streamlined mutation detection system: Multicolor post-PCR fluorescence labeling and single-strand conformational polymorphism analysis by capillary electrophoresis
A streamlined mutation detection system: Multicolor post-PCR fluorescence labeling and single-strand conformational polymorphism analysis by capillary electrophoresis
复制标题
DOI：
10.1101/gr.7.11.1094
复制
发表时间：
1997-11-01
期刊：
影响因子：
7
通讯作者：
Hayashi, K
中科院分区：
文献类型：
--
作者：
Inazuka, M;Wenz, HM;Hayashi, K
在遗传性疾病或癌症的研究中有效地利用人类基因组序列的知识在很大程度上取决于检测个体样本中突变的有效方法。我们在这里描述了一个简单而有效的突变扫描系统，其中PCR产物后标记两种不同的荧光染料在一个管中，并分析了自动毛细管电泳系统使用单链构象多态性（SSCP）条件（PLACE-SSCP）。通过适当使用内部对照DNA，精确评估参考和样品之间的电泳迁移率差异，然后统计判断突变的存在。使用一种电泳条件，在34个已知突变中的33个突变在三个不相关的序列背景的片段中检测到，最高可达741 bp，置信水平为
Effective use of knowledge of human genome sequences in studies of hereditary diseases or cancer heavily depends on efficient methods for detection of mutations in individual samples. We describe here a simple and efficient mutation scanning system in which PCR products are post-labeled with two different fluorescent dyes in one tube, and analyzed by an automated capillary electrophoresis system using single-strand conformation polymorphism (SSCP) conditions (PLACE-SSCP). With the appropriate use of an internal control DNA, differences in electrophoretic mobilities between a reference and samples are precisely evaluated, then the presence of mutations is statistically judged. Thirty-three of 34 known mutations in fragments of three unrelated sequence contexts up to 741 bp were detected using one electrophoresis condition at the confidence level of