ChIP-Atlas: SRX2416655

Antigen

library_strategy: ChIP-Seq
library_source: GENOMIC
library_selection: ChIP
library_construction_protocol: Cells were trypsinized and washed with PBS before fixing them with 1% paraformaldehyde. Fixed cells were used to prepare genomic DNA fargments using a sonicator. Sonicated fragments were immunoprecipitated using either anti-GATA2 or anti-GATA3 antibodies or non-specific IgG. Samples from three independent experiments were poooled and genomic libraries were sequenced in Illumina Genome Analyzer II and in Illumina HiSeq platforms to generate 35 bp single end reads. Libraries were prepared by Illumina. 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.