title: GSM1831385: WT + 2µ-URA3 vector with mtDNA_0; Saccharomyces cerevisiae; RNA-Seq
library_construction_protocol: Total RNA was extracted from 5 OD600 units of cells using the TRIzol reagent according to the manufacturer's instructions.  RNA was treated with RNase-free DNase I at a concentration of 1 U/µg to remove any remaining genomic DNA.  mRNAs were purified from 2µg total RNA using oligo (dT) magnetic beads and fragmented in fragmentation buffer using the TruSeq mRNA Sample Preparation Kit. Fragmented RNA was used for first-strand cDNA synthesis using a random hexameric primer, and the second strand was synthesized by using DNA polymerase I and RNase H.  Double-stranded cDNAs were purified with AMPure XP beads and eluted with re-suspension buffer, followed by 3'end adenine addition.  Sequencing adaptors were ligated to the fragments, and cDNA fragments were enriched by PCR amplification.  Enriched cDNA libraries were used for cluster generation and sequencing.  Paired-end sequencing of the cDNA libraries was performed using the Illumina Miseq platform.  All sequence data are PE 2x75 bp.  Image processing, base calling, and quality calue calculation were performed by the Illumina data processing pipeline (v1.5). High quality reads were saved in FASTQ format.