Transcriptome analysis of K + deficiency susceptible (PUSA362) and tolerant (PUSA372) cultivars under K + sufficient and deficient conditions.

Ten-days old desi Chickpea cultivars, K+ deficiency sensitive PUSA362 and tolerant PUSA372 were used for comparative transcriptome analysis under K+ and K- conditions. Total RNA was extracted from 100mg tissue sample using TRIZOL reagent (Ambion, life technologies USA) according to manufacturer's protocol. RNA samples were treated with DNaseI (Thermo Scientific) to remove any genomic DNA contamination, and samples were further purified using RNeasy MinElute Clean-up kit (QIAGEN). Quantification of RNA samples was done using nano-spectrophotometer (Nano Drop ONE Thermo Scientific) and RNA integrity was checked by loading the RNA samples on denaturing gel in 1X MOPS buffer. RNA quality was further ensured using an Agilent 2100 RNA Bioanalyzer (Agilent, USA). The library was prepared for RNA-seq experiment using the NEB Next Ultra II RNA Library Prep kit (NEB, Massachusetts, USA) using manufacturer instructions. The prepared library was quantified using Qubit 3 fluorometer (Thermofisher Scientific, Massachusetts, USA) using DNA HS assay kit (Thermofisher Scientific, Massachusetts, USA). The insert size of the library was assessed using 4200 TapeStation (Agilent Technologies, CA, USA). Prepared RNA library was sequenced in Illumina Novaseq 6000 at Nucleome Informatics, Hyderabad, India.