GSM2937033
GSM2937033: PRO-seq SNF5 rep1; Homo sapiens; OTHER
For ChIP-seq experiments, chromatin was clarified from sonicated nuclear preps and antibodies were added. For ATAC-seq, nuclei were extracted wtih non-ionic detergent in hypotonic buffer and treated with transposase. For RNA-seq, RNA was extracted by Trizol method, for PRO-seq, nuclei were extracted through hypotonic lysis with a low amount of non-ionic detergent. For ChIP-seq experiments, libraries were made using NEBnext Ultra II DNA library Prep and NEBNext Multiplex Oligos for Illumina, for ATAC-seq libraries were made using transposed DNA made Illumina Nextera DNA kit, NEBNext High-Fidelity 2x PCR Master Mix, and Nextera based primers ordered from IDT, for RNA-seq, libraries were made 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.
GEO
GSM2937033
GEO Accession
GSM2937033