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[A rapid and simple detection of DNA fragment with point mutation by capillary electrophoresis]
1The First Affiliated Hospital, West China University of Medical Sciences, Chengdu, Sichuan 610041.P.R.China. hungq@online.sc.cn
Objective:
To establish a rapid and simple method with high efficiency in detecting point mutation of genomic DNA.
Methods:
Four DNA fragments that were different from each other in only one based were amplified by using primers with artificial point mutation based on the sequence of exon 7 of p53 gene, and then were separated by capillary electrophoresis(CE). The neutral coated capillary and 4% linear polyacrylamide gel buffer were used, and the wave length of ultraviolet detector was 254 nm.
Results:
A homozygous 196 bp DNA fragment can be separated into one dsDNA peak and two ssDNA peaks within 25 minutes, and the heterozygous 196 bp DNA fragments that were made with mixed wild type and mutated DNA can be separated into one dsDNA peak and three ssDNA peaks. The three ssDNA fragments that differ in one nucleotide can be easily separated with good resolution.
Conclusion:
CE technique is rapid, sensitive, accurate and well reproducible. It is an efficient and reliable method for rapidly screening point mutation.