GSM2668147: dCcs1-SC_rep1; Saccharomyces cerevisiae; RNA-Seq
Total RNA was extracted by the hot acid-phenol method (Schmitt et al., 1990), and further purified by phenol-chloroform extraction. Following extraction and ethanol precipitation, total RNA was DNase-treated with Turbo DNase (ThermoFisher) according to the manufacturer's protocol. polyA+ RNA libraries were prepared using either the KAPA Stranded mRNA-seq library prep kit or the Illumina TruSeq Stranded mRNA Library Kit for NeoPrep, as indicated. Libraries were prepared according to the manufacturer's protocols. Both library preparation procedures first isolate polyA+ mRNAs using oligo-dT beads. Isolated polyA+ RNAs are then chemically fragmented, and random primers are used in the first cDNA reverse transcription step. Note that while both kits prepare "first strand" cDNA libraries, the strand-specific information was not used for data analysis in our study. However, that information can be extracted from the FASTQ files. Average library insert sizes were ~170 bp. High throughput sequencing was performed on Illumina’s HiSeq 4000 system, with single-end 50 bp insert reads, and dedicated index reads. Library prep: Illumina TruSeq Stranded mRNA Library Kit for NeoPrep
GEO Accession
GSM2668147