GSM1844611: GATA3 ChIP total thymocytes replicate 2; Homo sapiens; ChIP-Seq
DNA was purified by phenol/chloroform extraction. ChIP DNA were purified and 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 3â 'A' base for ligation of Illumina's adapters which have a single 3' overhanging 'T' base. After adapter ligation DNA was PCR amplified with Illumina primers for 25 cycles, and the product was selected for 200~300 bp (insert plus adaptor and PCR primer sequences) by gel electrophoresis and extraction (QIAex II, Qiagen). The purified DNA was subjected for a second round of PCR amplification for 10 cycles. Libraries were sequenced on the Genome Analyzer II following the manufacturer's protocols.