Instrument: Illumina HiSeq 2000
Strategy: RNA-Seq
Source: TRANSCRIPTOMIC
Selection: cDNA
Layout: SINGLE
Construction protocol: Entire cotyledons and juvenile leaves were harvested with sterile scissors and collected into RNase-free microfuge tubes, immediately snap-frozen in liquid nitrogen and stored at -80°C until RNA extraction. For each biological replicate (n = 3), a total of 30 and 20 pairs of cotyledons at Day 5 and Day 10, respectively, and 10 and two pairs of Leaf 1 and 2 at Day 10 and Day 16, respectively, were collected. Mature adult leaves (n = 3) were dissected to harvest the apical and basal thirds of the blade from the same Leaf 12 at Day 31, with the main vein in the basal section of leaves removed. These samples were also snap frozen in liquid nitrogen and stored at -80°C. Total RNA was extracted from samples using the RNeasy Plant Mini Kit (Qiagen) following the manufacturer's instructions, including two rounds of on-column gDNA elimination using DNase I digestion (Qiagen). The yield and purity of the extracted RNA was determined by Nanodrop (Thermo Fisher Scientific). At least 2.5 μg of total RNA from a single biological replicate was aliquoted into at least four technical replicates, to ensure at least three biological replicates from each single tissue type were processed for RNA-Seq. Total RNA samples were assessed and sequencing was performed by the Australian Genome Research Facility (AGRF, Melbourne). Sequencing libraries were constructed by Australian Genome Research Facility using the Illumina TruSeq stranded mRNA sample preparation protocol.