GSM1905479: yeast RNA-seq fast BioRep4; Saccharomyces cerevisiae; RNA-Seq
All solutions used were prepared with DEPC (diethylpyrocarbonate 0.1% v/v) treated water. Frozen sample aliquots were ground to a fine powder under liquid nitrogen(73).  Pestle and mortar were soaked in 10% bleach to destroy residual RNase activity and washed with diethylpyrocarbonate (DEPC) treated water. RNA was extracted using TriZol® reagent according to the methods of Hayes et al.(71) and the final concentration was measured prior to RNA sequencing using a NanoDrop system. Sequencing libraries were generated using the whole Transcriptome Library Preparation protocol provided with the SOLiD® Total RNA-Seq Kit (Life Technologies, Carlsbad, CA).  Briefly, rRNA depleted samples were fragmented using RNase III, and subsequently cleaned up using the RiboMinus™ Concentration Modules (Life technologies, Carlsbad, CA).  Fragmentation was assessed on a 2100 Bioanalyzer (Agilent Technologies, Palo Alto, CA) using the RNA picochip.  Fragmented RNAs were reverse transcribed and size selected on a denaturing polyacrylamide gel selecting for 150-250nt cDNA.  cDNA was then amplified and barcoded with SOLiD™ RNA barcoding Kit.  Samples were then purified using PureLink™ PCR Micro Kit (Life Technologies, Carlsbad, CA) and assessed on a 2100 Bioanalyzer (Agilent Technologies, Palo Alto, CA) using the High Sensitivity DNA chip.
GEO Accession
GSM1905479