GSM8730037: spn1 141-305 Rep1; Saccharomyces cerevisiae; RNA-Seq
RNA was isolated from yeast expressing either wild-type Spn1, spn1K192N, or spn1141-305 in triplicate 10-ml cultures by hot-acid phenol extraction as previously described (Iyer and Struhl 1996). Cells were grown in YPD at 30°C to mid-exponential phase (OD600 ~ 1.0), harvested, and each sample was spiked to obtain a final concentration of 1 S. pombe to 10 S. cerevisae cells. Messenger RNA was purified from total RNA using poly-T oligo-attached magnetic beads. After fragmentation, the first strand cDNA was synthesized using random hexamer primers followed by the second strand cDNA synthesis. The library was ready after end repair, A-tailing, adapter ligation, size selection, amplification, and purification. The library was checked with Qubit and real-time PCR for quantification and bioanalyzer for size distribution detection. Quantified libraries were pooled and sequenced on Illumina platforms.