GSM1577756: RNASeq IL2 rep1; Homo sapiens; RNA-Seq
For RNA-Seq,  RNA from 5 donors was pooled, and 1 μg of the pooled RNA was used to synthesize cDNA 220-400 bp fragments were isolated using 2% E-Gel (Invitrogen), ends were repaired, adaptor was added using T4 DNA ligase (New England Biolabs), and amplified for 17 cycles using PE 1.0 and PE 2.0 primers (Illumina) and Phusion High Fidelity PCR Master Mix (New England Biolabs). For ChIP-Seq, Chromatin from 10-20 million cells was sonicated into 250-500 bp fragments, DNA ends were repaired using polynucleotide kinase and Klenow enzyme, and treated with Taq polymerase to generate a protruding 3' “A” nucleotide. DNA fragments of ~250-450 bp were purified from agarose gels and used for cluster generation and sequencing. Antibody against STAT5B was used for immunoprecipitation. PCR products were barcoded (indexed) and sequenced on Illumina HiSeq 2000 platform.
GEO Accession
GSM1577756