title: GSM4478121: 2601: KSHV (-) tumor cells; Mus musculus; MBD-Seq
library_construction_protocol: Genomic DNA was isolated using DNeasy Blood & Tissue Kit (QIAGEN) and sheared by sonication to fragments of ~500bp. Methylated DNA enrichment protocol: The sheared DNA (1µg) was added to 10 µl MBD-Bead slurry (MethylMiner DNA Enrichment Kit, Invitrogen, Carlsbad, CA) and incubated on a rotating mixer for 1 hr, as described previously [49]. The DNA fragments were eluted into distinct subpopulations based on the degree of methylation; non-captured fraction (NC, representing un-methylated DNA fragments), 450 mM NaCl (representing partially methylated DNA fragments) and 2000 mM NaCl (representing methylated DNA fragments). The fractions were then ethanol precipitated and re-suspended in H2O. MBD2 enriched methylated DNA fractions were subjected to library preparation with NEBNext® Ultra™ II DNA library prep kit for Illumina (NEB #E7645) at the next generation genomic center in the Azrieli Faculty of Medicine, Bar-Ilan University.
attributes.tag: GEO Accession
attributes.value: GSM4478121