Figure 1
From: Improved reduced representation bisulfite sequencing for epigenomic profiling of clinical samples
Key laboratory steps in RRBS. The isolated DNA from the samples are digested by two restriction enzymes (TaqαI and MspI). The fragments are then end-repaired and ligated with adapters. The ligated DNA are size-selected and repetitive sequences are removed. PCR amplification is performed after bisulfite conversion. Illumina GAIIx system is used for high-throughput sequencing. Newly added bases (marked in bold in step 2) are always unmethylated and thus discarded in the downstream analysis for CpG methylation.