Establtshing the standard control of MS-PCR (methylation - specific PCR) reaction to determine methylation of brca1 gene promoter in breast cancer samples

Abstract

In this study, genomic DNA from the PC3 cancer cell line and the recombinant plasmid containing the methylated BRCA1 sequence have been isolated from transformants. The linear plasmid DNA and bisulfite treated PC3 DNA which carry the methylated BRCA1 sequences have been used as templates for PCR with the MS- PCR primers. The results have shown that the MS-PCR has been capable of detecting 6 copies of the methylated BRCA1 sequence per 10000 copies of the non-methylated BRCA1 sequence from linear plasmid and 2 copies from PC3 genomic DNA. These results have been absolutely consistent with that of the previous reports, which has indicated that linear recombinant plasmid and genomic DNA extracted from breast cancer cell lines are suitable for establishing the standard control of the MS-PCR method. The authors have suggested that the linear plasmid DNA should be used as a positive control in the MS-PCR to determine the methylated BRCA1 sequence in breast cancer samples.