Figure 1.

Overview of the MBD-seq workflow.

Step 1) Genomic DNA is randomly fragmented with ultrasonication. Step 2) Methylated fragments are captured by a protein with high affinity for double-stranded DNA harbouring methylated CpGs. Unmethylated DNA fragments are washed away. Step 3) The captured portion of the methylome is eluted. Step 4) The methylation enriched fraction (the elute) is used to generate barcoded sequencing libraries. Step 5) Barcoded libraries are pooled in equal molarities. 6) The library pool is sequenced and aligned to the reference genome. Step 7) The aligned reads are analysed with suitable software such as RaMWAS that was specifically developed for large-scale methyome-wide association studies.