Genome-wide analysis of DNA methylated status of CpG islands with the modified AIMS technology / 解放军医学杂志

ABSTRACT

Objective To conduct the global assessment of methylated DNA status of CpG islands. Methods The modified AIMS (amplification of inter-methylated sites) technology was adopted, which mainly included 3 groups of isoschizomers (methylation-sensitive and methylation-insensitive restricted enzymes). The comparison analysis of methylome among 3 concentrations (6%, 8%, 10%) of polyacrylamide with 8M urea denaturing sequencing gel and the digestion effect among 3 groups of isoschizomers were carried out. The results of personal molecular imager FX system and autoradiography were also compared. Results Genome-wide detection of methylome of CpG islands might be the technical basis for the subsequent analysis. The resolving power and scope bands in 6% polyacrylamide denaturing sequencing gel were superior to those in gels with the other two concentrations. 3 groups of isoschizomers could meet the need of genome-wide detection of methylome of CpG islands. Personal molecular imager FX system and autoradiography could be used together, and they compensated each other. Conclusions The optimized AIMS technology is a kind of high-throughput method to analyze methylome, which is simple, specific, and easy to handle. The 3 groups of isoschizomers can cover most of CpG islands for genome-wide detection and get the ideal results. Only a small quantity of genome DNA is enough to meet the need of clinical detection.