title: GSM3682098: [ESI+] Diploid mRNA-seq, replicate 1; Saccharomyces cerevisiae; RNA-Seq
library_construction_protocol: Total RNA was extracted using a standard hot acid phenol Total RNA was extracted from diploid yeast strains grown to mid-exponential phase (OD600 ~0.5) using a standard hot acid phenol protocol followed by chloroform extraction and ethanol precipitation. Afterwards, RNA was resuspended in DEPC-treated water and quantified on a NanoDrop spectrophotometer. Further quality control and ribosomal RNA evaluation was performed by analysis on an Agilent Bioanalyzer 2100 Nano RNA assay at the Protein and Nucleic Acid Facility (Stanford). For library generation, ERCC spike-in controls (Invitrogen) were added, and total RNA was reverse-transcribed by on-bead polyA-tail priming with Superscript IV (Invitrogen), and then fragmented on a Covaris E220 to a size range of 200–400 bp. Fragmented cDNA was end-repaired and A-tailed, and dual-index sequencing adaptors were ligated using the NEBNext Ultra II DNA Library Prep Kit. Prepared libraries were sequenced on the Illumina HiSeq 4000 platform at the Stanford Functional Genomics Facility using paired-end reads.
attributes.tag: GEO Accession
attributes.value: GSM3682098