title: GSM2258040: RNAseq_Spt5-T0-untagged_II; Schizosaccharomyces pombe; RNA-Seq
library_construction_protocol: RNA was extracted using the hot phenol method (PMID: 12854975 ) For RNA-seq libraries, 200 µg of total RNA from each strain was spiked-in with 10 µl of 1:100 diluted ERCC control RNA(). Then, poly(A)+ RNA was enriched by two rounds of Dynabeads Oligo (dT)25 (Invitrogen) purification, followed by alkaline fragmentation and size selection of fragments  40-70 nucleotides. Size-selected RNA was dephosphorylated and ligated to an RNA linker at the 3’ end. First-strand cDNA was synthesized using a primer with sequence complementary to the 3’-linker and an additional adapter sequence, using Superscript III reverse transcriptase (Invitrogen). cDNA was circularized using CircLigase (Epicentre). The library was amplified with 8-14 cycles of PCR, using primers specific for the adaptor sequence and the products were gel purified. Library size distributions and concentrations were determined using an Agilent Bioanalyzer. All strains were done in duplicate. RNA-seq libraries were sequenced on the Illumina HiSeq platform at the Tufts University Core Facility.
attributes.tag: GEO Accession
attributes.value: GSM2258040