title: GSM3444230: IBC_H3K27ac_Std_PAT-ChIP; Homo sapiens; ChIP-Seq
library_construction_protocol: Chromatin was extracted from FFPE sections of 10 µm thickness using the standard PAT-ChIP protocol ChIP DNA was blunt-ended and phosphorylated, and a single 'A' nucleotide was added to the 3' ends of the fragments in preparation for ligation to adapters that have a single-base 'T' overhang using enzymes and reagents from NEB (New England Biolabs). The ligation products were purified and size-selected by Agencourt AMPure XP beads (Beckman Coulter). Purified DNA was PCR-amplified with PfuUltra II Fusion HS DNA Polymerase (Agilent Technologies) to enrich for fragments that have adapters on both ends. The final purified product was then quantitatively and qualitatively checked on Bioanalyzer 2100 (Agilent Technologies). Libraries with distinct adapter indexes were multiplexed (1/5 libraries per lane) and after cluster generation on FlowCell were sequenced for 50 bases in the single read mode on a HiSeq 2000 sequencer (Illumina Inc.).