GSM6429721: AEP1 (WT) + preQ1-L1, IP, rep2 (Expt1); Schizosaccharomyces pombe; RIP-Seq
Total RNA was isolated from 50 OD of cells. Cells were harvested and after addition of 1 mL of phenol, glass beads and vigorous shaking for 5 min, samples were centrifuged at 20.000 g for 5 min to clear the cell debris. Equal volume of phenol/chloroform/isoamylalcohol was added to the aqueous phase and centrifuged at 20.000 g for 5 min. After mixing the upper phase with an equal volume of chloroform followed by centrifugation at 20.000 g for 5 min, the RNA was precipitated. Library preparation of immunoprecipitated RNAs for deep sequencing was performed using the NEBNext Small RNA Library Prep Set for Illumina (Multiplex Compatible; New England Biolabs) according to the manufacturer's protocols.