GSM5345458: RPE.Ctrl.H3K36me3.G1.rep2; Homo sapiens; ChIP-Seq
Lysates were clarified from sonicated nuclei and histone-DNA complexes were isolated with antibody. ChIP sequencing libraries were prepped using the TruSeq ChIP Sample Preparation kit (Illumina) following the supplier's protocol with slight modifications. 1 ng or 35 ml of DNA was used for library preparation. After ligating the adapters, fragments were amplified with 13 cycles of PCR (“Purify Ligation Products” section was skipped). The PCR products were purified using a Double-Sided SPRI approach: AMPure was added to 0.6X SPRI concentration (30 ml of AMPure is added to 50 ml of PCR reaction) and incubated for 10 minutes at RT before being placed on the magnet. Beads were discarded and supernatant was incubated with 0.85X SPRI concentration (12 ml of AMPure is added) and incubated for 10 minutes at RT before being placed on the magnet. Liquid was removed before performing two washes with 450 ml fresh 70% EtOH. DNA was eluted with 15 ml H2O. Library length distribution were assessed by TapeStation (Agilent) and quantified with Qubit (ThermoFisher). Libraries were single-end sequenced (75 cycles) using a NextSeq500 (Illumina).
GEO Accession
GSM5345458