Memory CD8+ T cells, triggered through CD3/CD28 signaling for 4 hr
Memory CD8+ T cells
anti-p300 (Santa Cruz, #sc-585X)
Sequenced DNA Library
Histone-DNA complexes were isolated with a p300-specific antibody Libraries were prepared according to Illumina's instructions accompanying the TruSeq_DNA_SamplePrep Kit. 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. Libraries were sequenced on the Genome Analyzer following the manufacturer's protocols.