Miltenyi Biotec naive CD4+ T cell magnetic bead isolation
Sequenced DNA Library
library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
Lysates were clarified from sonicated nuclei and histone-DNA complexes were isolated with antibody. DNA was end-repaired using a combination of T4 polynucleotide kinase and T4 DNA polymerase. The blunt phosphorylated ends were treated with Klenow fragment (3' to 5' exo-) and dATP to yield a single 3' A to blunt DNA for ligation of Illumina adapters with a single 3' T overhang. After adapter ligation DNA was PCR amplified with Illumina adapters for 19 cycles and library fragments of ~250 bp (insert plus adapter and PCR primer) were selected by bead purification. The purified DNA was captured on an Illumina flow cell for cluster generation. Libraries were sequenced on the Illumina Hiseq 2500 following the manufacturer's instructions.