GSM5088036: wt biological replicate 3; Saccharomyces cerevisiae; RNA-Seq
Total RNA was extracted from yeast cells using the Bead-beat Total RNA Mini kit (A&A Biotechnology, catalog no. 031-25BB ) and purified using Clean-Up RNA Concentrator (A&A Biotechnology, catalog no. 039-25C) mRNA was enriched using oligo(dT) beads. The mRNA was fragmented randomly, and then cDNA was synthesized using an mRNA template and random hexamers primer, after which a custom second-strand synthesis buffer (Illumina), dNTPs, RNase H, and DNA Pol I were added to initiate second-strand synthesis. After a series of terminal repair and ligation of sequencing adaptors, the double-stranded cDNA library was completed through size selection and PCR enrichment.