GSM1483713: BY_RM_18min_R1; Saccharomyces cerevisiae; RNA-Seq
The collected yeast cells from each time point were stored at -80°C for no more than 2 days before we extracted total RNA from the cells using a standard phenol-chloroform preparation. We used a TruSeq RNA Sample Prep v2 Kit (Illumina) to create a sequencing library from the total RNA collected for each decay rate time course time point from each replicate. Per the protocol for the kit, we isolated mRNA from the total RNA using two rounds of poly(A) selection, then fragmented the isolated mRNA into approximately 150 base pair fragments, and finally, used random hexamer primers to produce cDNA. We created barcoded sequencing libraries from the cDNA from each sample and, in an effort to minimize technical variation between the data acquired from different decay rate time points, all samples from all replicates were sequenced in the same lane on an Illumina HiSeq 2000 (50 bp paired-end reads).
GEO Accession
GSM1483713