GSM2656049: internal SPEN_HS_43_30'_rec4h; Mus musculus; ChIP-Seq
The ChIP assay was carried out according to the protocol of a ChIP kit from Upstate Biotechnology (Lake Placid, NY) using protein A-sepharose beads (Amersham). Standard crosslinking procedure was used. For each IP reaction 48-50 µg of chromatin sonicated to 100-500 bp fragments and 5 µg of anti-SPEN polyclonal antibodies, or IgG from rabbit serum (Sigma) as a negative control were used. Immunoprecipitated DNA was additionally digested by RNase A (in a final concentration 70 μg/ml, 30 minutes, 37°C) before Proteinase K treatment and organic solvents extraction. Glycogen (70 μg/ml) was added at the precipitation step. DNA was dissolved in H2O. In each experimental point five to six ChIP replicates were collected and combined in one sample before DNA sequencing. Sequencing libraries were generated using KAPA library preparation kit (for Illumina) KK8201. Template amplification and cluster generation were performed using the TruSeq SBS Kit.
GEO Accession
GSM2656049