GSM9557291: PS_CAMHB_rep2; Pseudomonas syringae pv. tomato str. DC3000; RNA-Seq
Total RNA was isolated and prepared from cell pellets using reagents from Zymo Research and silica-coated magnetic beads (Luna Nanotech) accroding to the vendor's manual. Concentrations and quality of the RNA were assessed using an Agilent TapeStation and Nanodrop, respectively. For each sample, 1 ug was used as input for the standard RiboRid procedure. RNA-seq libraries were prepared from 5 uL of the rRNA-subtracted RNA using KAPA RNA HyperPrep Kit (Roche) according to the manufacturer's protocol with volumes reduced by half. The final PCR amplification step was modified by incorporating SYBR and using a qPCR instrument. Samples were removed from the thermal cycler just before entering the plateau phase of amplification to avoid biases being potentially introduced by reagent depletion. The quality and concentrations of the libraries were checked with an Agilent TapeStation and Qubit instrument, respectively. Pooled libraries were then run on an AVITI instrument (Element Biosciences).