GSM1439699: Tet2_flox_AML1_ETO_p10; Mus musculus; Bisulfite-Seq
Immature myeloid cells (GFP+, CD11b-, Gr1-, CD16/32+) were sorted from AML1-ETO cultures. Genomic DNA was isolated using the AllPrep DNA/RNA micro kit (Qiagen). Enhanced Reduced Representation Bisulfite Sequencing (eRRBS) libraries were generated as previously described (Akalin, A., et al (2012), PLoS Genet. 8, e1002781) with the exception that end-repair and adaptor ligation were performed using the NEBNext® DNA library Sample Prep Master kit (New England Biolabs). In brief, ~1ug of genomic DNA was digested with MspI enzyme overnight followed by phenol/chloroform extraction. After library generation using pre-annealed methylcytosine-containing illumina adaptors (TruSeq® DNA sample prep kit, Illumina) and size-selection (150-250bp and 250-400bp), the ligated fragments were treated with bisulfite (EZ DNA methylation kit, Zymo) to convert unmethylated cytosines. Finally, the converted fragments were PCR-amplified (15x PCR cycles), pooled and sequenced on an Illumina Hiseq2000.
GEO Accession
GSM1439699