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[应用毛细管电泳-PCR-SSCP分析检测p53基因点突变]

[Detection of point mutation in p53 gene by capillary electrophoresis-PCR-SSCP analysis].

作者信息

Liu Ji, Jing Huie, Ru Qinhua, Deng Yingping, Luo Guoan, Huang Qian

机构信息

Department of Biochemistry and Molecular Biology, West China School of Preclinical and Forensic Medicine, Sichuan University, Chengdu 610041, China.

出版信息

Sichuan Da Xue Xue Bao Yi Xue Ban. 2003 Jul;34(3):476-9.

Abstract

OBJECTIVE

To screen the point mutation of p53 gene rapidly by capillary electrophoresis (CE).

METHODS

A simple capillary electrophoresis system for the strand conformational polymorphism (SSCP) analysis of genomic DNA was developed by choosing commercially available capillary and gel buffer. This CE-SSCP system was applied to the analysis of twenty PCR products amplified from the exon 7 of the p53 gene of tissue specimens from patients with colon cancer.

RESULTS

Five of them (Ca4, Ca6, Ca7, Ca8, Ca14) were found to have mutation within 25 minutes, while the results of the SSCP analysis by PAGE-silver staining techniques only detected mutation of four samples. And sequencing of these PCR products proved that the results of CE-SSCP were correct.

CONCLUSION

This simple, accurate and less expensive method could be used in the field of medical research and in the clinical DNA diagnosis of human cancers and other diseases.

摘要

目的

通过毛细管电泳(CE)快速筛选p53基因的点突变。

方法

选用市售毛细管和凝胶缓冲液,建立了一种用于基因组DNA链构象多态性(SSCP)分析的简单毛细管电泳系统。该CE-SSCP系统应用于分析从结肠癌患者组织标本的p53基因第7外显子扩增的20个PCR产物。

结果

其中5个(Ca4、Ca6、Ca7、Ca8、Ca14)在25分钟内被发现有突变,而通过PAGE-银染技术进行的SSCP分析仅检测到4个样本的突变。对这些PCR产物进行测序证明CE-SSCP的结果是正确的。

结论

这种简单、准确且成本较低的方法可用于医学研究领域以及人类癌症和其他疾病的临床DNA诊断。

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