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目的 建立一种新的非同位素 SSCP分析方法。方法 常规提取人类基因组 DNA作为模板 ,PCR扩增 nm 2 3- H 1基因 ,变性 PCR产物 ,6 %非变性聚丙烯酰胺凝胶 SSCP电泳 ,银染后分析结果。结果 改进了目前 PCR- SSCP分析中应用同位素标记的检测方法 ,采用银染方法 ,建立起简单迅速的非同位素 PCR- SSCP分析方法和适宜条件。并将这种方法成功地应用于肿瘤转移抑制基因点突变的检测。结论 非同位素聚合酶链反应 -单链构象多态 (Nonisotopic PCR- SSCP)分析是一种迅速、敏感地检测基因顺序中突变的单链 DNA聚丙烯酰胺凝胶电泳技术。
Objective To establish a new non-isotopic SSCP analysis method. Methods Routine extraction of human genomic DNA as a template, PCR amplification of nm 2 3-H 1 gene, denatured PCR product, SSCP electrophoresis on 6 % non-denaturing polyacrylamide gel, and analysis results after silver staining. RESULTS: The method of isotope label detection in the current PCR-SSCP analysis was improved, and a simple and rapid non-isotopic PCR-SSCP analysis method and suitable conditions were established using the silver staining method. This method was successfully applied to the detection of point mutations in tumor metastasis suppressor genes. Conclusion Nonisotopic polymerase chain reaction-single strand conformational polymorphism (Nonisotopic PCR-SSCP) is a single-stranded DNA polyacrylamide gel electrophoresis technique for the rapid and sensitive detection of mutations in gene sequences.