GSM4626705: O DNA methylation replicate 1; Mus musculus; Bisulfite-Seq
FACS-isolated MuSCs were snap-frozen. Genomic DNA was extracted using the QIAamp Micro DNA extraction kit (Qiagen). DNA (150 ng) was sheared with a Bioruptor 300 (Diagenode) and then end-repaired and A-tailed with the NEBNext DNA Library Prep kit (NEB). Methylated Illumina adapters were ligated with LigaFast (Promega), and size selection was performed with AMPure XP beads (Beckman Coulter). Bisulfite conversion was performed with the EZ DNA Methylation Direct kit (Zymo) and amplified with 14 cycles with PfuTurbo Cx Hotstart DNA Polymerase (Agilent) followed by final size selection with the AMPure XP beads. Libraries underwent paired-end 101-bp sequencing at the Stanford Genome Sequencing Service Center with an Illumina HiSeq 2000 at low cluster density with PhiX spike-in to a depth of 115-200 million reads.
GEO Accession
GSM4626705