GSM1981430: ada2 null mutant (YSJ4); [Candida] glabrata; RNA-Seq
The 20 ml cultures were immediately added to 30 ml pre-cooled methanol to make final 60% methanol in order to stop the cellular processes and RNase activity. Cells were centrifuge at -4°C and the total RNAs were extracted using the RiboPureTM-Yeast Kit (Ambion). Total RNAs were extracted and treated with DNase to remove DNA contamination. The mRNAs were enriched by oligo(dT) magnetic beads and then broken into short fragments (~ 200 bp). The first strand of cDNA was synthesized using random hexamer primer, while second strand was synthesized using DNA polymerase I. The double strand cDNA was further purified with magnetic beads. End reparation and 3' end single nucleotide A (adenine) addition was performed. Sequencing adaptors were ligated to these fragments and fragments were enriched by PCR. During the QC step, Agilent 2001 bioanalyzer and ABI StepOnePlus Realtime PCR System were used to qualify of the sample library. The library products were sequencing via Illumina HiSeqTM 2000.
GEO Accession
GSM1981430