GSM5463094: RNAseq H2B WT Replicate1; Homo sapiens; RNA-Seq
MDA-MB-231 cells expressing FLAG-tagged wild-type or mutant histones were seeded in 6-well plates.  The cells were collected and total RNA was isolated using the RNeasy kit (Qiagen, 74136) according to the manufacturer's instructions. The total RNA was then enriched for polyA+ RNA using Dynabeads Oligo(dT)25 (Life Technologies, 61002).  The polyA+ RNA was then used to generate strand-specific RNA-seq libraries as described previously (Zhong et al., 2011).  The RNA-seq libraries were subjected to QC analyses (i.e., number of PCR cycles required to amplify each library, the final library yield, and the size distribution of final library DNA fragments) and sequenced using an Illumina NextSeq 500 to an average depth of ~40 million reads total per condition.  Two independent biological replicates were used.