Fig. 1

Schema of BisPCR² method for targeted bisulfite sequencing. DNA sequencing libraries are prepared by bisulfite conversion of genomic DNA followed by two rounds of PCR for target enrichment (PCR#1) and subsequent sample barcoding (PCR#2). Partial adapter overhangs are added to target enrichment primers to permit simplified library preparation by PCR. PCR#1 amplicons are pooled prior to the PCR#2 reaction for each biological sample. Due to the presence of the unique barcodes, all PCR#2 amplicons can be pooled for a single next-generation sequencing run.