Plant sample from Coffea arabica x Coffea canephora 0 hai
Leaves were inoculated with spores of H. vastatrix race XXXIII and sampled at 0, 12, 24, 96 hours after inoculation (hai) and 17 days after inoculation (dai). Then total RNA was extracted with RNeasy Plant Mini Kit (Qiagen), according to the manufacturer's instructions, and cDNA synthesis were performed using the kit Mint-2 cDNA Synthesis (Evrogen). After this step, the samples were normalized with kit TRIMMER – cDNA Normalization (Evrogen), to reduce the abundant transcripts. RNA sequencing were performed using the Miseq platform (Illumina) at University of North Carolina Chape Hill, NC, USA, using the protocol kit TruSeq DNA Sample Preparation (Illumina). This sequencing produced 2x250pb paired-end reads for each genotype and time sampled (Freitas, 2015) and these 10 libraries produced (5 time points for each of the 2 genotypes) were sequenced 10 MiSeq runs, each run comprising all libraries in a multiplex reaction, using barcodes for each library.