Instrument: Illumina HiSeq 2000
Strategy: RNA-Seq
Source: TRANSCRIPTOMIC
Selection: cDNA
Layout: PAIRED
Construction protocol: Total RNA from 200 mg of ground material was extracted using TRIzol (Invitrogen, Karlsruhe, Germany) following the manufacturer’s instructions. Prior RNA extraction, plant materials harvested at 3, 7 and 14 dpi from the same independent biological experiment were pooled together. As control total RNA from 3 independent biological replicates of seven-day-old S. vermifera grown in MYP medium was used. RNA samples were additionally precipitated with ethanol. In brief, 1/10 volume of 3 M NaOAc and 3 volumes of ethanol were added to RNA solution. After incubation at - 20°C overnight, the RNA pellet was centrifuged at 13000 rpm for 30 min and washed once with 70% ethanol (diluted in DEPC ddH2O) and spin down for 10 min. The RNA pellet was then air-dried and resuspended in RNase-free water with a final concentration of 1 μg/μl. Purity and quantity of RNA samples were measured using the NanoDrop ND-1000 spectrophotometer (NanoDrop Technologies, Wilmington, DE, USA) and Agilent RNA 6000 Nano Kit and Agilent 2100 Bioanalyzer following the manufacturer’s protocol (Agilent, Santa Clara, USA). cDNA libraries were prepared for sequencing using standard Illumina protocols by IGA Technology Services (Udine , Italy)