GSM6997978: Sample 7: 1110 GS cells, normoxia, RRBS; Canis lupus; Bisulfite-Seq
DNA was isolated using Zymo Research Quick DNA Mini-prep Plus kit. 40 ng of starting input genomic DNA was digested with 30 units of MspI (NEB). Fragments were ligated to pre-annealed adapters containing 5'-methyl-cytosine instead of cytosine according to Illumina's specified guidelines. Adaptor-ligated fragments ≥50 bp in size were recovered using the DNA Clean & Concentrator™-5 (Cat#: D4003). The fragments were then bisulfite-treated using the EZ DNA Methylation-Lightning™ Kit (Cat#: D5030). Preparative-scale PCR was performed and the resulting products were purified with DNA Clean & Concentrator™-5 (Cat#: D4003) for sequencing on an Illumina NovaSeq 6000 platform (150 bp PE reads). 