GSM6856755
Chromatin Immunoprecipitation experiments were carried out in M121224 cell lines using Active Motif ChIP-IT High sensitivity kit as per manufacturer's instructions. for H3K27Ac 10ug of chromatin and 3ug of H3K27ac (ab4729, Abcam) antibodies were used. Immunoprecipitation was carried out overnight at 4C with rotation. Libraries were prepared using the NEBNext® Ultra DNA Library Prep for Illumina® kit (ref. E7370) according to the manufacturer's protocol. Briefly, input and ChIP enriched DNA were subjected to end repair and addition of “A” bases to 3′ ends, ligation of adapters and USER excision. All purification steps were performed using AgenCourt AMPure XP beads (ref. A63882, Beckman Coulter). Library amplification was performed by PCR using NEBNext® Multiplex Oligos for Illumina (Index Primers Set 1, ref. E7335), (Index Primers Set 2, ref. E7500), (Index Primers Set 3, ref. E7710) and (Index Primers Set 4, ref. E7730). Final libraries were analyzed using Agilent Bioanalyzer or Fragment analyzer High Sensitivity assay (ref. 5067-4626 or ref. DNF-474) to estimate the quantity and check size distribution, and were then quantified by qPCR using the KAPA Library Quantification Kit (ref. KK4835, KapaBiosystems) prior to amplification with Illumina's cBot