GSM8231027: yeast, noIAA, rep1; Saccharomyces cerevisiae; RNA-Seq
Total RNA was extracted using the NucleoSpin RNA kit (Macherey-Nagel) following manufacturer's instructions. Ribosomal RNA was removed from total RNA, followed by ethanol precipitation. After fragmentation, the first strand cDNA was synthesized using random hexamer primers. During the second strand cDNA synthesis, dUTPs were replaced with dTTPs in the reaction buffer. The directional library was obtained following end repair, A-tailing, adapter ligation, size selection, USER enzyme digestion, amplification, and purification.