title: GSM3439862: Dib1-6h-rep1; Saccharomyces cerevisiae; RNA-Seq
library_construction_protocol: Total RNA was extracted using RiboPure Yeast RNA extraction kit (Ambion), and purified using Rneasy column purification kit (Qiagen), including a Dnase digestion step. Quality and integrity of the RNA samples were assessed using a 2200 TapeStation (Agilent Technologies) and then libraries generated using the TruSeq® Stranded mRNA assay (Illumina, Inc.) according to the manufacturer's protocol. Briefly, total RNA (0.1-4ug) was used as input material from which polyadenylated mRNA was purified using poly-T, oligo-attached, magnetic beads. The mRNA was then fragmented using divalent cations under elevated temperature and then reverse transcribed into first strand cDNA using random primers. Second strand cDNA was then synthesised using DNA Polymerase I and RNase H. Following a single 'A' base addition, adapters were ligated to the cDNA fragments, and the products then purified and enriched by PCR to create the final cDNA library. Adapter indices were used to multiplex libraries, which were pooled prior to paired-end sequencing (151:151 plus indices) on an Illumina NextSeq500 platform using a NextSeq 500/550 High Output Kit. Finally, the output data was demultiplexed (allowing one mismatch) and BCL-to-Fastq conversion performed using Illumina's bcl2fastq software, version 2.17.1.14
attributes.tag: GEO Accession
attributes.value: GSM3439862