T0 Transcriptome of the taxol-producing endophytic fungus Cladosporium cladosporioides MD2
About 60 ¦Ìg of total RNA at a concentration of ¡Ý 600 ng/¦Ìl, OD260/280=1.8~2.2 and RNA integrity number (RIN) ¡Ý 7.0 were used for construction of cDNA sequencing library using a TruSeqTM RNA Sample Preparation Kit (Illumina, San Diego, California, USA). Briefly, poly-A mRNA samples were isolated from the total RNA using oligo (dT) attached magnetic beads and then interrupted into 200-500 bp fragments by fragmentation buffer consisted of metallic cations. Subsequently, the fragmented mRNA were used for the synthesis of first-strand cDNA with random hexamer primers and reverse transcriptase, and then further synthesized the second-strand cDNA using DNA polymerase I and RNase H. The cDNA products were purified with QIAquick PCR Purification Kit (Qiagen, California, USA), end-repaired and A-tailed using End Repair Mix (Qiagen), and then ligated with sequencing index adapters (Illumina). The ligated products isolated by gel electrophoresis to select suitable fragments were amplified to generate the final cDNA libraries, that was sequenced using Illumina HiSeq? 2500 sequencing platform.