Post-conversion targeted capture of modified cytosines in mammalian and plant genomes. Homo sapiens

We present a capture-based approach for bisulphite-converted DNA that allows interrogation of pre-defined genomic locations, allowing quantitative and qualitative assessments of 5-methylcytosine (5mC) and 5-hydroxymethylcytosine (5hmC) at CG dinucleotides and in non-CG contexts (CHG, CHH) in mammalian and plant genomes. We show the technique works robustly and reproducibly using as little as 750 ng of starting DNA, with results correlating well with whole genome bisulphite sequencing data, and demonstrating that human DNA can be tested in samples contaminated with microbial DNA. This targeting approach will allow cell type-specific designs to maximize the value of 5mC and 5hmC sequencing.