title: GSM3593629: PRO-seq WT rep2; Homo sapiens; OTHER
library_construction_protocol: Chromatin was clarified from sonicated cells and protein-DNA complexes were isolated with antibody. ChIP libraries were prepared according to New England Biolabs  instructions accompanying the NEBNext Ultar II DNA Library Prep Kit for Illumina (Part# E7645S). After adapter ligation DNA was PCR amplified with NEBNext Multiplex Oligos for Illumina (Part# E7335S and E7500S) for 16 cycles (HA ChIP) or 15 cycles (WDR5 ChIP). For RNA-seq, libraries  were prepared using Poly-A selected at Genewiz using their standard methods, and for PRO-Seq, libraries were made using RNA that was made through a biotin-run on reaction.Biotin-RNA was base hydrolyzed, and extracted. 3'-adaptors were ligated to ends and 5'caps were removed, repaired, and then 5'-adaptors were ligated as well. Purified and intact biotin-RNA containing adaptors was used in a reverase transcriptase reaction to generate cDNA and cDNA was used to amplify full library using NEB High Fidelity Phusion polymerase.RNA libraries were prepared using Poly-A selected RNA.
attributes.tag: GEO Accession
attributes.value: GSM3593629