Seqcap ez pure capture bead kit

The TAB-SeqCapEpi protocol requires the TAB-Seq kit (WiseGene, IL, USA) and the SeqCap Epi Choice Enrichment library together with the compatible library prepation reagents (KAPA Lib Prep kit, SeqCap Adapter kit, SeqCap EZ Hybridization and Wash kit, SeqCap Epi Accessory kit, SeqCap HE-Oligo kit and SeqCap EZ Pure Capture Bead kit) (Roche Nimblegene, WI, USA). Sonicated genomic DNA samples (1–1.5 μg) were split into BS- and TAB-aliquots (1/3 and 2/3 of the input volume, respectively). The TAB-aliquots were treated with βGT and then oxidized with the mTet1 enzyme from the TAB-Seq kit. Then both BS- and TAB-aliquots were subjected to the SeqCap Epi Enrichment System protocol v1.0 with the exception that up to six aliquots (corresponding to three input gDNA samples) were pooled prior to hybrid capture. The full TAB-SeqCapEpi workflow is described in Supplementary Text S4 (Supplementary File 1).

FP gene capture was performed using the SeqCap EZ Hybridization and Wash Kit (Roche, Basel, Switzerland), SeqCap EZ Accessory Kit v2 (Roche), SeqCap HE-Oligo Kit (Roche), and SeqCap EZ Pure Capture Bead Kit (Roche) following the manufacturer’s instructions for SeqCap EZ library SR (Roche), with minor modifications. Briefly, Biotin-labeled FP-gene fragments were used as hybridization probes, instead of the SeqCap EZ library (Roche). Equal amounts of all libraries (in total, 1 µg) were mixed with 135 ng of Biotin-labeled FP-gene fragments and were hybridized at 46°C for 72 h. Other procedures were performed in accordance with the manufacturer’s instructions.