HIGH-THROUGHPUT SEQUENCING DETECTION METHOD FOR METHYLATED CPG ISLANDS

摘要

A high-throughput sequencing method for detecting methylated CpG islands comprises: (a) treating a DNA sample with a modifying agent to form the modified DNA wherein cytosine bases but not 5'-methyl-cytosine bases of the DNA sample are modified to uracil bases; (b) providing Primer A and DNA polymerase to amplify the modified DNA and obtain the semi-amplicon which is able to anchor Adapter Primer C at one end; (c) amplifying the semi-amplicon using Primer B and polymerase to obtain the full-amplicon which enrich the methylated CpG islands in the DNA sample and is able to anchor Adapter Primer C at one end and Adapter Primer D at the other end; (d) amplifying the full-amplicon using Adapter Primer C and D; and (e) sequencing the full-amplicon. The process enriches and amplifies the methylated CpG islands from a converted genomic DNA sample by means of a high-CpG-density oligonucleotide primer, simultaneously anchoring the adapter sequences to the amplicon for high-throughput sequencing.