GSM1624639: Control ST-HSC; Mus musculus; Bisulfite-Seq
As previously published in Gu et al. Nat Protoc. 2011 Apr;6(4):468-81 with slight modifications Enhanced RRBS (eRRBS) was performed on 10 ng DNA purified from LT-HSC, ST-HSC, or MPP isolated from aged male Vav-IDH1-KI mice and control littermates (WT) (n=6/group) using a FACS Aria instrument (Beckton Dickenson). For ligation, methylated adapters from TruSeq Kit (Illumina) were diluted 1:4 and 1.5 μl were used in the reaction. After bisulfite conversion and before fragment selection, gels were cut between 150-400 bp for fragment size selection. Final libraries were purified using Agencourt AMpure XP magnetic beads (Beckman Coulter) according to the manufacturer’s protocol. Libraries were sequenced at the Princess Margaret Genomics Centre using a HiSeq2000 instrument. Five libraries per lane were multiplexed and 30% PhiX was added to increase library complexity.