GSM2475253: RNAseq.EB_d2.KO.rep1; Mus musculus; RNA-Seq
Total RNA was isolated with TRIzol (Life Technologies) and reverse transcribed using Superscript III and random hexamers (Life Technologies) to synthesize the 1st strand. Second strand was synthesized with dUTP to generate strand asymmetry using DNA Pol I (NEB, M0209L) and the E. coli ligase (Enzymatics, L6090L). Libraries were repaired to generate blunt 5' P ends using 'EndIT DNA End Repair kit' (Epicentre), 3' A overhang was generated using 'Klenow Fragment (3´→ 5´ exo-)' (NEB).  Illumina barcodes were ligated using 'Rapid T4 DNA Ligase' (Enzymatics). Next libraries were size selected using agarose gel extraction method.  After around 13 cycles of amplification, libraries were cleaned, quantified and pooled.
GEO Accession
GSM2475253