GSM2337660: Stage 0 uninfected; Solanum lycopersicum; miRNA-Seq
Root tissue of equal weight of each disease development stage was processed for RNA isolation using TRI reagent (Sigma-Aldrich, St. Louis, MO, U.S.A.) as per manufacturer’s instructions. The integrity of RNA was checked on 1.2% agarose gel and quantification was done through optical density measurement using nanovue (GE Healthcare Bio-sciences, Uppsala, Sweden). In total 11 small RNA libraries were prepared including five infected disease development stages and their corresponding uninfected tissue of same age and a library of five-week old uninfected root tissue of susceptible tomato cv. Small RNA libraries from root tissue of each stage were prepared independently using Illumina small RNA sample preparation kit (Illumina, San Diego, CA, U.S.A.) according to manufacturer’s instructions. Briefly, an equal amount of total RNA of each stage was used for sequential ligation of 3’and 5’adapter using T4 RNA ligase. Adapter ligated small RNA was reverse transcribed using SuperScript II reverse transcriptase (Invitrogen, Carlsbad, CA, U.S.A.) and 3’ adapter specific RT-primer. cDNA prepared was amplified and product of approximately 147 bp was eluted through 6% polyacrylamide gel. The quality and quantity of cDNA libraries prepared were determined through Agilent 2100 bioanalyzer (Agilent Technologies, Santa Clara, CA, U.S.A.). High-throughput sequencing of libraries was performed using Illumina Hiseq 2000 system at Institute of Genomics and Integrative Biology, Delhi, India, according to manufacturer’s instructions.
GEO Accession
GSM2337660