GSM5311327: rpb1-FSP1; Saccharomyces cerevisiae; RNA-Seq
RNA was extracted from cell pellets using an acid phenol procedure followed by pelleting in CsCl2, resuspension, ethanol precipitation, and purification by Qiagen Rneasy mini kit. Total RNA samples (100-500 ng) were hybridized with Ribo-Zero Gold to substantially deplete cytoplasmic and mitochondrial rRNA from the samples.  Stranded RNA sequencing libraries were prepared as described using the Illumina TruSeq Stranded Total RNA Library Prep Gold kit (20020598) with TruSeq RNA UD Indexes (20022371).  Purified libraries were qualified on an Agilent Technologies 2200 TapeStation using a D1000 ScreenTape assay (cat# 5067-5582 and 5067-5583).  The molarity of adapter-modified molecules was defined by quantitative PCR using the Kapa Biosystems Kapa Library Quant Kit (cat#KK4824).  Individual libraries were normalized to 1.30 nM in preparation for Illumina sequence analysis. Each library was sequenced on two lanes of an Illumina SP Flow Cell on a Novaseq 6000 instrument as 2 x 50 bp reads.