title: GSM5104890: hEBw5 P300 rep1A; Homo sapiens; ChIP-Seq
library_construction_protocol: Organoids were rinsed in medium, homogonized in a 2ml Kontes Glass Co. dounce and immediately crosslinked after. Cells were lysed, chromatin was sheared to ~500bp fragments and chromatin was extracted using antibody-coated beads. DNA was purified using RNase cocktail enzyme mix, proteinase K, two phenol:chloroform:iso-amyl-alcohol(25:24:1) and 2 chloroform:iso-amyl-alcohol(24:1) extractions followed by DNA precipitation in ethanol and Zymo PCR purification columns. TruSeq DNA Sample Preparation kit (Illumina), following the Low Throughput guidelines with the following additions: Size selections were performed before and after amplification on an E-gel Safe Imager (Invitrogen) using 2% E-gel SizeSelect gels (Invitrogen). The ChIP-DNA fraction of 300-400 bp in size (including adapters) was isolated and purified. For adapter ligations, 1 ul instead of 2.5 ul of DNA Adapter Index was used. ChIP-seq 100bp paired end reads on a Illumina HiSeq 2000 device, trimmed to 85bp for mapping