title: GSM1182898: dSP22c: rRNA-depleted total RNA; Mus musculus; RNA-Seq
library_construction_protocol: RNA greater than 200 nt from d7 ES cells and MEFs was harvested using the mirVana RNA extraction kit (Ambion), cleared of ribosomal RNA (Ribozero, Epicentre) and sheared to a median size of 200 nt, followed by library construction. RNA fragments were end-repaired (T4 polynucleotide kinase, NEB), ligated to 3' linker (T4 RNA ligase2 trunc., NEB), reverse transcribed (Superscript III, Invitrogen) and ligated to a 3’linker on the previous 5' end. This single-stranded cDNA library was then amplified (Phusion, NEB) using custom oligos matching the Illumina TruSeq PCR primers.