RNA-Seq of Phaffia rhodozyma: OEGTF5-28h
Total RNA was extracted and mRNA was enriched by magnetic beads with Oligo(dT). The mRNA was broken into short fragments and used as a template to synthesize single-stranded cDNA with random hexamer primers, then synthesized double-stranded cDNA which was purified with a kit. The purified double-stranded cDNA was subjected to end repair, added poly(A) tail and connected to a sequencing adapter, then the fragment size was selected, and finally amplified by PCR. After the constructed library was qualified by Agilent 2100 Bioanalyzer, Illumina HiSeq X Ten was used for sequencing to generate 125 bp or 150 bp paired data.