Sudden transition from jin1 to eds8-1, passage 4
Pools were made from 20 symptomatic plants per condition at passages 1, 4, 7, and 10.  These were frozen with liquid N2 and stored at 80 C until they were homogenized into a fine powder.  For RNA-seq, total RNA was extracted from 70 to 90 mg of infected and healthy plants using the GeneJET Plant RNA Purification kit (Thermo Fisher Scientific, Waltham, MA, USA) according to the manufacturers instructions.  RNA quality was assessed using a NanoDrop One (Thermo Fisher Scientific) and agarose gel electrophoresis, while integrity and purity were evaluated with a Bioanalyzer 2100 (Agilent Technologies, Santa Clara, CA, USA).  Library preparation and Illumina sequencing were performed by Novogene Europe using a NovaSeq 6000 platform, the Lnc-stranded mRNA-Seq library method, ribosomal RNA depletion, directional library preparation, 150 bp paired-end reads, and 6 Gb of raw data per sample.  Novogene performed a quality check of the libraries using a Qubit 4 Fluorometer (Thermo Fisher Scientific), qPCR for quantification, and a Bioanalyzer for size distribution detection.
SUB15074160
Sudden_jin1-eds8_4