GSM1315606: CBP_null1_EtOH; Mus musculus; ChIP-Seq
Cell lysates were sonicated (5x 10sec @ 15micron amplitude, MSE Soniprep 150) and immunoprecipitated using CBP antibody (MEFs lacking CBP provide control for antibody specificity) library prepared according to Illumina protocol "Preparing Samples for ChIP Sequencing of DNA" (2007).   Briefly, DNA was end-repaired using a combination of T4 DNA polymerase, E. coli DNA Pol I large fragment (Klenow polymerase) and T4 polynucleotide kinase. The blunt, phosphorylated ends were treated with Klenow fragment (32 to 52 exo minus) and dATP to yield a protruding 3- 'A' base for ligation of Illumina's adapters which have a single 'T' base overhang at the 3’ end. After adapter ligation DNA was PCR amplified with Illumina primers for 15 cycles and library fragments of ~250 bp (insert plus adaptor and PCR primer sequences) were band isolated from an agarose gel. The purified DNA was captured on an Illumina flow cell for cluster generation.
GEO Accession
GSM1315606