Smoking 2, Smokehouse 1, Time 4
Grape berries were pulverized under liquid nitrogen. Total RNA was extracted from three biological replicates using a modified DEPC-CTAB protocol (Reinth et al. 2014). RNA was quality-checked on an agarose gel and was quantified using a Nanodrop 2000 spectrophotometer (Thermo Fisher Scientific, Waltham, MA, USA). Following quality validation and quantification using a Qubit Fluorometer (Thermo Fisher Scientific) and Agilent Bioanalyzer (Santa Clara, CA, USA), cDNA libraries were prepared from the RNA and sequenced by the WSU Genomics Core (Spokane, WA) on an Illumina HiSeq 2500platform as 2x100 bp paired-end reads.