GSM2770698: Pto_flg22_#1-1; Pseudomonas syringae pv. tomato str. DC3000; RNA-Seq
RNA was extracted using the TRIzolTM reagent (Thermo Fisher Scientific) and the Direct-zolTM RNA Miniprep kit (Zymo Research). Purified RNA was treated with 10 U of RNase-free DNase I (Roche Applied Science), after which RNA was purified a second time with the Direct-zolTM RNA Miniprep kit. Non-organellar 18S and 28S rRNAs, and poly(A) mRNAs were depleted using the MICROBEnrichTM kit (Thermo Fisher Scientific). RNA-Seq libraries were prepared using the Ovation Arabidopsis RNA-Seq system 1–16 (NuGEN) with two modifications to the manufacturer's protocol. First strand cDNA synthesis used only the first strand primer random mix (the oligo dT primer mix was omitted), while on strand selection II, custom insert dependent adapter cleavage probes (InDA-C; currently referred to as AnyDeplete) with specificity to highly abundant A. thaliana chloroplast and nuclear transcripts (361 probes) and Pto DC3000 rRNAs (65 probes) (1 µL of each 2 µM probe mixture was used) were added to the mixture of A. thaliana cytoplasmic, chloroplast, and mitochondrial rRNA custom probes that are included with the kit.
GEO Accession
GSM2770698