RNA-Seq of Geosmithia pallida
Theextracted total RNAs amounting to 5.0 g were resuspended in RNase-free water and stored at80C until use. The extracted RNA samples were used for cDNA synthesis. The poly (A)mRNA was isolated using oligo-dT beads (Qiagen). All mRNA was broken into short fragments(200 nt) by adding a fragmentation buffer. First-strand cDNA was generated using randomhexamer-primed reverse transcription, followed by the synthesis of the second-strandcDNA using RNase H and DNA polymerase I. The cDNA fragments were purified using aQIAquick PCR extraction kit. The purified fragments were then washed with EB buffer for endpreparation poly (A) addition and ligated to sequencing adapters. The cDNA fragments(200  25 bp) were purified and enriched via PCR to construct the final cDNA library followingagarose gel electrophoresis and cDNA extraction from gels. The cDNA library was sequencedon the Illumina sequencing platform (Illumina HiSeq 2000) using the single-end paired-end(PE) technology within a single run.