GSM2199580: Input_DNA; Caenorhabditis elegans; ChIP-Seq
FLAG::CEBP-1-DNA complexes were isolated with anti-FLAG antibody. Both ChIPed DNA and input genomic DNA were ligated to specific adaptors and amplified by barcode primers. 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 (3' to 5' exo minus) and dATP to yield a protruding 3- 'A' base for ligation of adapters which have a single 'T' base overhang at the 3’ end. After adapter ligation DNA was PCR amplified with primers for 12 (1st) or 18 (2nd) cycles with DNA polymerase (KAPA) and library fragments of ~250 bp (insert plus adaptor and PCR primer sequences) were band isolated from an agarose gel. The purified DNA was quantified using Qubit. Libraries were sequenced on the Genome Analyzer following the manufacturer's protocols.
GEO
GSM2199580