iPSC_Input4_InputH3K27ac_R01L01S01
For control input chromatin, 5-15 x 10^6 formaldehyde crosslinked cells were lysed and sonicated in 110ul of SDS Lysis Buffer (0.5% SDS, 50mM Tris-HCl pH 8.0, 20mM EDTA, 1x cOmplete Protease Inhibitor Cocktail (Sigma)) using Covaris E220 Focused-ultrasonicators (Covaris) for 14 cycles, 1 min per cycle, duty cycle 5. Input chromatin was then reverse crosslinked by incubation for 15 min at 65C with rotation and subsequent incubation with 5 ul RNAse (Sigma) for 1h at 37C and Proteinase K Solution (20 mg per mL, Thermo Fisher Scientific) for 1h at 55C. After reverse crosslinking, samples were purified with 2X Agencourt AMPure XP DNA beads (Beckman Coulter), eluted in 30 ul of H2O and Qubit (Thermo Scientific) quantified. Libraries were generated using KAPA Hyper Prep Kit (KAPA Biosystems) and KAPA Real Time Library Amplification Kit (KAPA Biosystems) following manufacturers manual. Libraries were barcoded using TruSeq RNA Indexes (Illumina), size selected for 300 bp to 500 bp, and sequenced with either 100 bp paired-end or 150 bp paired-end reads on an Illumina HiSeq 4000.