RNAseq of Ascochyta rabiei infecting chickpea 12hai replicate 2 Read 1
Total RNA was isolated from  A. rabiei infecting chickpea leaves 12 hai. Three independent replicates were performed.RNA was extracted using TRISure.  Potential residual genomic DNA was removed using RQ1 RNase-Free. RNA was further purified using the RNeasy Plant Mini Kit. RNA-seq libraries were generated from 5?g of total RNA by first capturing the mRNA using Oligo dT(25) beads. The purified mRNA was then randomly fragmented in a Zn2+ solution and Illumina sequencing adapters were ligated to the RNA. cDNA was then generated using a p5 oligonucleotide for priming and the library was finally amplified with 10 cycles of PCR. To distinguish A. rabiei reads from those originating from chickpea, reads were mapped against our own preliminary A. rabiei genome assembly and against the chickpea transcriptome reported by Hiremath et al (2001) using the TopHat2.0.3 software. Only reads mapping to the A. rabiei genome but not to the chickpea transcriptome were retained