RNA-Seq of Brettanomyces bruxellensis: response to sulfite stress
Samples containing 5 mL of culture were mixed with 10 mL of cold RNAlater (Life Technologies, Melbourne, Australia). Tubes were then centrifuged for 10 min at 4000 rpm and 4C and, after discarding the supernatant, the cell pellets were frozen and stored at -80C. RNA extraction was performed using the RiboPure-Yeast Kit as described by the manufacturer (Life Technologies, Melbourne, Australia). Extracted RNA was quantified using the Quant-iT RiboGreen RNA Assay Kit (Life Technologies, Melbourne, Australia). RNA sequencing (RNASeq) libraries were prepared using the Illumina Truseq mRNA kit and sequenced using single-ended 100 bp chemistry (AGRF, Australia and The Ramaciotti Centre for Genomics, Australia).