Strand-specific RNA-Seq of Aspergillus terreus MUCL38669: Submerged culture under the influence of exogenous butyrolactone I.
The submerged culture of Aspergillus terreus MUCL38669, where exogenous butyrolactone I had been added at 120 hours p.i., was sampled at 24, 48, 96, 120, 144 and 216 hours p.i. Total RNA was separately extracted from each sample and pooled prior to the strand-specific double stranded cDNA library production. The library was prepared as follows: mRNA was obtained using oligo-dT magnetic beads followed by random fragmentation. The first-strand cDNA was synthesised using reverse transcriptase with random hexamer primers. In the second-strand cDNA synthesis dNTPs were used with an exception of dTTPs being replaced by dUTPs. Subsequently, Illumina mRNA preparation protocol was used with one exception: The dU-containing second strand was digested with Uracil-N-Glycosylase prior to the library amplification in order to obtain strand-specific paired-end sequencing reads.