title: GSM3370093: BSGX001-thi2-2; Saccharomyces cerevisiae; RNA-Seq
library_construction_protocol: The cells in each samples were collected by centrifuging at 5000 rpm, 4°C, for 5 minutes. Then the pellets were rapidly freezed in liquid nitrogen and stored at −80°C. UNIQ-10 Trizol RNA Purification Kit (Sangon Biotech, China) was used to extract total RNA. The total RNA was extracted and fragmented, the DNA was digested with DNase I, enriching the eukaryotic mRNA with the magnetic beads with Oligo (dT) (if it is a prokaryote, then remove the rRNA from the reagent into the next). Add interrupting agent in Thermomixer to break the mRNA into short fragments. The cDNA is synthesized using the short fragments mRNA as template, then the two-stranded cDNA was synthesized by make two chain synthesis reaction system, the Short fragments are purified and resolved by the kit, the 3 'end of cDNA is connected with the base "A" and is connected with the connector, then suitable fragments were selected and the PCR amplification is performed. The constructed library is analyzed with the Agilent 2100 Bioanalyzer and ABI StepOnePlus Real-Time PCR System,then sequenced using Illumina HiSeqTM 2000.
attributes.tag: GEO Accession
attributes.value: GSM3370093