GSM8061322: WGBS of P60 M+N+ (eGFP+) sorted beta-cells from female mice - Rep 3; Mus musculus; Bisulfite-Seq
Mice were euthanized either two days (P2) or 2-months after birth. Their islets were isolated, dissociated into single cells, and used for FACS cell sorting based on the expression of eYFP and Apple fluorescent proteins. Dead cells were excluded using DAPI staining. Collected cells ere immediately processed for genomic DNA isolation using Zymo kit D4011. Tagmentation-based WGBS (T-WGBS) libraries were prepared very similarly to previously reported methods (Adey and Shendure 2012, Barnentt et al. 2020, Wang et al., 2013). Genomic DNA from purified beta-cell subtypes were extracted using a kit from Zymo. For each stage/sex, at least three batches of cells from FACS (4-7 mice for each batch, at least 30,000 cells) were combined, yielding 100-150 ng DNA. Library concentrations and size distributions were evaluated using an Agilent 2200 TapeStation with a D5000 screentape. T-WGBS DNA libraries were sequenced at Vanderbilt University's genomics research core, VANTAGE, using 2×150bp paired-end reads on the Illumina NovSeq 6000, obtaining approximately 300-500 million reads per library.