GSM1253106: C.M1M2.1; Saccharomyces cerevisiae; RNA-Seq
We extracted total RNA from samples using Ambion RiboPure-Yeast Kit. We purified mRNA from total RNA using Ambion Micro PolyPurist Kit, and reversed transcribed mRNA into cDNA using polyA primers. We used T4 DNA Polymerase (NEB), T4 Polynucleotide Kinase (NEB), and Taq Polymerase (NEB) for end repair of the sheered cDNA.  We used T4 ligase (NEB) to ligate index adapters to the cDNA . We pooled each sample to an equal concentration and gel extracted 250 – 350 bp of ligated cDNA and used PCR to amplify the selected fragment to a final concentration of 1 nM.  We submitted our final cDNA library to GTAC at Washington University for sequencing on two Illumina HiSeq lanes. At a run concentration of 6 pM
GEO Accession
GSM1253106