title: Whole genome DNA methylation of undifferentiated spermatogonia of postnatal day 14 (P14) wild-type and Spocd1-K464A (does not interact with TPR) homozygous mutant mice
design: Whole genome DNA methylation of undifferentiated spermatogonia of postnatal day 14 (P14) wild-type and Spocd1-K464A (does not interact with TPR) homozygous mutant mice
library_construction_protocol: FACS-purified P14 undifferentiated spermatogonia (DAPI -ve, CD45 -ve, CD51-ve, c-Kit -ve, CD9 +ve) DNA from spermatogonia was isolated by Proteinase K digest (10 mM Tris-HCl pH 8, 5 mM EDTA , 1% SDS, 0.3 M Na-acetate, 0.2 mg/ml proteinase K) overnight, followed by 2x phenol/chloroform/isoamylalcohol (25:24:1, buffered at pH 8, Sigma-Aldrich) extraction and 2x chloroform extraction. The DNA was precipitated at -20˚C after addition of 1/10 volume 3M Na-acetate, 10µg linear acrylamide (Invitrogen) and 1 volume of isopropanol, washed 2x with 70% ethanol, and solubilized in 5mM Tris pH8. Methyl-seq libraries were prepared using the NEBnext Enzymatic Methyl-seq kit (NEB E7120) according to the manufacturer's instructions
identifiers.namespace: European Bioinformatics Institute
identifiers.id: E-MTAB-14862:WT_1_p