F-SSCP: fluorescence-based polymerase chain reaction-single-strand conformation polymorphism (PCR-SSCP) analysis.
F-SSCP: fluorescence-based polymerase chain reaction-single-strand conformation polymorphism (PCR-SSCP) analysis.
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F-SSCP：基于荧光的聚合酶链反应-单链构象多态性（PCR-SSCP）分析。
DOI：
10.1101/gr.2.1.10
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发表时间：
1992
期刊：
影响因子：
--
通讯作者：
K. Hayashi
中科院分区：
文献类型：
--
作者：
R. Makino;H. Yazyu;Y. Kishimoto;T. Sekiya;K. Hayashi
开发了一种基于荧光的聚合酶链反应-单链构象多态性 (PCR-SSCP) 分析方法 F-SSCP，其中使用荧光引物通过 PCR 扩增目标序列。 The amplified products are then heat-denatured and applied to a water-jacket controlled gel in an automated DNA sequencer.分离的链在凝胶底部被检测为激光激发荧光，突变被检测为荧光图中峰位置的移动。该系统不涉及放射性，并且电泳条件比以前依靠环境空气冷却来保持凝胶恒温的系统控制得更严格。输出数据的性质允许直接定量解释，因此可以轻松估计两个或多个等位基因的混合物中每个等位基因的相对丰度。据报道，F-SSCP 用于检测肿瘤组织中 p53 突变和杂合性缺失。
A fluorescence-based method for polymerase chain reaction-single-strand conformation polymorphism (PCR-SSCP) analysis, F-SSCP, was developed in which the target sequence is amplified by the PCR using fluorescent primers. The amplified products are then heat-denatured and applied to a water-jacket controlled gel in an automated DNA sequencer. The separated strands are detected as laser-excited fluorescence at the bottom of the gel, and mutations are detected as shifts in the position of the peaks in the fluorogram. The system does not involve radioactivity, and the conditions of electrophoresis are more strictly controlled than in the previous system, which relied on ambient air-cooling to maintain the gel at a constant temperature. The nature of the output data allows direct quantitative interpretation, and so the relative abundance of each allele in a mixture of two or more alleles can easily be estimated. The application of F-SSCP for detection of mutations and loss of heterozygosities of p53 in tumor tissues is reported.