PCR-RFLP中Chelex-100制备DNA模板的方法建立及其条件优化

A reliable, economical and large-scale method was established for molecular epidemiology and population genetics investigations. According to the SNP rs679620 (-Lys45Glu-) in exon 2 of the MMP3 gene, a pair of primer was designed for PCR. Using the Chelex-100 Method to extract genomic DNA from a micro amount of human whole blood, the genotypes was obtained by PCR-RFLP. The optimized condition is: 10μL of human whole blood with 200μL of 5 % Chelex-100 solution in a sample tube was immersed in a 56℃ water bath for 30min, then in a 100℃ water bath for 8mim, taking a sample of the supernatant after centrifugation. By PCR amplification and enzyme digestion, the target DNA belt(s) was clearly shown by agarose gel electrophoresis. The Chelex-100 method for extracting human genomic DNA is a simple, fast, reliable and economical method. It is especially suitable for analysis of human genotypes by large scale DNA extraction in a massive population.