Plant sample from Coffea canephora
Total RNA was isolated from 60 mg of leaf tissue (between six and seven circular explants) using Plant/Fungi Total RNA Purification Kit with DNase treatment (Norgene, Cat. 25800 and 25710 respectively) according to the manufacturer's instructions, in a final volume of 25 L. To avoid degradation, all steps were carried out at low temperature. RNA concentration and integrity were determined using a NanoDropTM 2000 spectrophotometer (Thermo Scientific) and 1.2 % agarose gel electrophoresis. For RNA preservation, the RNAstable Tube Kit (Sigma, 93221-001) was used according to the manufacturer's instructions. For the preparation and sequencing of the libraries, the service provided by the sequencing company Novogene (https://www.novogene.com/us-en/) was used. RNA-Seq libraries were sequenced on an Illumina Platform to a sequencing depth of at least 20 million read pairs (150 bp paired-end sequencing) per sample