GSM1695655: Tomato_control; Solanum lycopersicum; ncRNA-Seq
Total RNA from the leaf samples (at 21-days of post inoculation) was extracted using the mirVana miRNA isolation kit (Ambion, Austin, TX, USA), with modifications. Briefly, 100 mg of leaf sample was ground with 800 µl of Lysis/Binding buffer. Then, 80 µl of miRNA homogenate was added and the resulting solution was mixed by vortexing, and then placed on ice for 10 minutes. Acid phenol-chloroform extraction was then performed. Samples were treated with the DNase RQ1 (Promega, MA) for 1 h at 37 ºC and then the chloroform extraction was performed. RNA integrity was examined in a 2100 Bioanalyzer (Agilent Technologies, California, USA). The small RNA of 15- to 50-nt was purified from the total RNA samples and then subjected to small RNA library preparation using NEBNext Multiplex Small RNA Library Prep Set for Illumina according to manufacturers instructions (New England Biolabs). RNA libraries obtained for the RNA extracted from the control plants, PSTVd-M inoculated plants and PSTVd-I inoculated products were processed simultaneously in the Illumina Hi-Seq platform using the multiplex strategy.
GEO Accession
GSM1695655