GSM3560179: MYBB_dsmo_G2- rep.1; Homo sapiens; ChIP-Seq
Cells (107) were cross-linking was achieved using 1% methanol-free formaldehyde in RPMI 1640 for 10 min at room temperature. Cells were ressupended in 1% SDS in TE-PMSF buffer and sonicated (Bioruptor; Diagenode, Seraing, Belgium) at 4oC for 15 cycles (30 s ON/30 s OFF).Immunoprecipitation was done with 5ug anti-phosphoT487-BMYB (ab76009, ABCAM) or without any antibody (Input) and 20 uL of protein G coupled to magnetic beads (Thermo-Fisher) in 100 uL total volume, incubated overnight at 4oC. The cross-linking reversal was done with 0.5 mg proteinase K and reversion buffer for 30 min at 65oC. To produce the libraries were used the TrueSeq DNA kit (Illumina; San Diego, CA, USA), according to the manufacturer´s instructions. The sequencing in a HiSeq2000 equipment. 50 bp fragments were sequenced by the single-end method, with the standart 7 bps index reading and V3 chemistry.
GEO Accession
GSM3560179