GSM2794879: WT_rep2; Saccharomyces cerevisiae; RNA-Seq
Total RNA from yeast cells was isolated by RNeasy midi kit (Qiagen).The mRNA was purified from total RNA by Dynal oligo(dT) beads (Invitrogen). The first and second strand cDNA was synthesized by SuperScript III CellsDirect™ cDNA Synthesis Kit (Invitrogen) and SuperScript® Double-Stranded cDNA Synthesis Kit (Invitrogen), respectively; the resulting double strand cDNAs were followed by DNA repair and end-polishing (blunt-end) with End-It DNA End-Repair Kit (Epicentre), purified with QIAquick PCR Purification Kit(Qiagen), and dA-tailed utilizing the 3´-5´ exo-Klenow Fragment (NEB),the resulting purified fragments were ligated to adaptor oligo mix (Illumine) using Quick T4 DNA ligase (NEB). The 200-500 bp ligation products were recovered from a 2% (w/v) agarose gel using Qiagen gel extraction kit. Then followed by a PCR amplification with Solexa primers using KAPA HiFiTMHotStart kit. The 250-400 bp amplified products were purified again from a 2% agarose gel and used directly for high throughput sequencing.
GEO Accession
GSM2794879