GSM1260179: 979B3_Aid+/+ iPS cells_Nanog-GFP_p6_cultured on gelatin; Mus musculus; MBD-Seq
Cells were lysed with genome buffer (8.8 mM Tris-HCl pH8.0, 135 mM NaCl, 8.8 mM EDTA, 1% SDS, 200 μg/mL protenase K) and treated with RNase. Subsequently genomic DNA was purified with Phenol-Chloroform Isoamyl Alcohol (PCI) DNA extraction. Genomic DNA was sonicated to 100-300 bp fragment with Covaris E210 (Covaris). Methylated DNA was enriched with EpiXplore Methylated DNA Enrichment Kit (Takara) according to manufacturer’s protocol. Multiplexed MBD-seq libraries were prepared from 10 ng of metylated DNA fragment using NEBNext ChIP-seq Library Prep Master Mix (New England BioLabs). For sequencing using GAIIx, cluster generation was performed using TruSeq SR Cluster Kit v2. Each lane of flow cells contained one sample. Sequencing was performed by a single-read run mode with total 76 cycles, including 75-bp read and one cycle for phasing.
GEO Accession
GSM1260179