GSM2124966: JS167_eif3e; Schizosaccharomyces pombe; RNA-Seq
Total RNA was extracted by phase separation using RNA-BeeTM (Amsbio) and chloroform, and precipitated in isopropanol. RNA was further purified using the RNeasy Mini Kit (Qiagen), performing an on-column DNase digestion. The quality of total RNA was assessed by the Agilent Bioanalyzer (Agilent Technologies). Total RNA samples were treated with RiboMinus Eukaryotic kit (Invitrogen) to deplete rRNA. The rRNA-depleted sample was subjected to enzymatic fragmentation using RNase III from the SOLiD Total RNA-Seq kit. The purified fragmented RNA was then ligated with SOLiD adaptors on both ends and served as templates for cDNA synthesis. The cDNA libraries were constructed and purified twice using AMPure beads to ensure complete capture and size-selection of desired cDNA. The cDNA was PCR amplified and sequenced on the SOLiD 4 platform.
GEO Accession
GSM2124966