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[Detection of point mutation in p53 gene by capillary electrophoresis-PCR-SSCP analysis]
1Department of Biochemistry and Molecular Biology, West China School of Preclinical and Forensic Medicine, Sichuan University, Chengdu 610041, China.
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.