GSM3289967: EP1_HDAC11; Homo sapiens; ATAC-seq
Chromatin was extracted and processed for Tn5-mediated tagmentation and adapter incorporation, according to the manufacturer's protocol (Nextera DNA sample preparation kit, Illumina®) at 37 °C for 30 minutes. Reduced-cycle amplification was carried out in presence of compatible indexed sequencing adapters. Total cellular RNA was purified using the Qiagen RNAeasy Mini kit and samples with RNA integrity number ≥ 7 were further processed. total RNA was denatured at 65°C for 5 minutes, cooled on ice, purified and incubated at 80°C for two minutes. The eluted mRNA was fragmented at 94°C for 8 min and converted to double stranded cDNA, end repaired, A-tailed, and ligated with indexed adaptors. The quality of the libraries was assessed by DNA-based fluorometric assay (Thermo Fisher ScientificTM) and automated capillary electrophoresis (Agilent Technologies, Inc.). Up to 3 samples per lane were pooled and run on a HiSeq2500 Illumina sequencer with a paired-end read of 50bp. The quality of the libraries was also assessed by RNA-based fluorometric assay (Thermo Fisher ScientificTM) and automated capillary electrophoresis (Agilent Technologies, Inc.).