GSM1661791: RNA Pol II ChIP on A771726-treated cells.; Homo sapiens; ChIP-Seq
Sample information curated by ChIP-Atlas
Cell type Class
Attributes by original data submitter
A375 melanoma cell line
25 μM A771726
Sequenced DNA Library
DNA was sonicated and chromatin immunoprecipitation was performed with either HEXIM1 (Abcam), CDK9 (Santa Cruz) or RNA Pol II (Santa Cruz) antibodies. DNA was then de-cross-linked from pulled down protein and purified by phenol:chloroform:isoamyl alcohol extraction. The NEBNext multiplex oligos for Illumina Kit (NEB) was used to make the samples suitable for multiplexing. The PCR reaction was run for 18 cycles. The indexed libraries were purified on a 2% agarose gel, and were cut out between 150 and 350 bp. The concentration of the isolated libraries was estimated with a high-sensitivity DNA chip from Agilent according to manufacturer’s protocol and libraries were mixed in equal quantities and sequenced on Illumina Hi-Seq2000. Index sequences from multiplexed primers were used to identify treatment group and reads were demultiplexed using a perl script.