GSM1924870: Input for ChIP-Seq in mouse neural stem cells; Mus musculus; ChIP-Seq
ChIP was performed using 107 mouse neural stem cells per reaction. Cells were dissociated by treatment with Accutase and cross-linked in 1% (vol/vol) formaldehyde for 10 min at RT with rotation. Then, 0.125 M glycine was used to quench the cross-linking reaction. Cells were pelleted and washed with ice-cold PBS. Next, nuclei were isolated and a Bioruptor sonicator (Diagenode) was used to shear chromatin DNA. Either 5 ml of OLIG2 antibody (AB9610, Chemicon) or 5 ml of normal rabbit IgG (#2729, Cell Signaling) was added to Dynabeads Protein A (Invitrogen) beads and incubated for 3 h at 4 °C with rotation. Then, the Dynabeads-antibody complexes were incubated with sheared chromatin DNA overnight at 4 °C. After immunoprecipitation, the precipitated complex was treated with RNase A and Proteinase K, and incubated at 65 °C overnight to reverse crosslinks. The ChIP-Seq library was constructed by using DNA SMART ChIP-Seq Kit according to the manufacturer's instructions (Clontech) and was sequenced on the Illumina HiSeq 2000 Sequencer.
GEO Accession
GSM1924870