GSM1543653: polyA+ RNA from the cytoplasmic fraction-E2, rep 1; Homo sapiens; RNA-Seq
RNA-seq libraries were generated from two biological replicates for each condition.  The total RNA was extracted using the PARIS kit (Ambion).  After purification, 4000 ng of Total RNA was subjected to enrichment of polyA+ RNA using Dynabeads Oligo(dT)25 (Invitrogen) as described previously (Zhong et al., 2011). Strand-specific RNA-seq libraries were prepared from the polyA+ RNA according to the “deoxyuridine triphosphate (dUTP)” method as described previously (Zhong et al., 2011). The RNA-seq libraries were sequenced using an Illumina HiSeq 2000 as follows: (1) the polyA+ RNA-seq libraries from E2- or control-treated fractionated RNA were sequenced using paired-end methodology with a length of 100 nt (PE100) (two replicates each) and (2) the polyA+ RNA-seq library from untreated unfractionated RNA was sequenced using single-end methodology with a length of 50 nt (SE100) (one replicate).