RNA-seq of filamentous fungi: Talaromyces pinophilus
The mycelia of Talaromyces pinophilus were cultured with common fungal medium for different time, and harvested for RNA extracting use the fungal RNA extracting kit following the manufacturers protocol. Oligo(dT)-attached magnetic beads were used to purified mRNA. Purified mRNA was fragmented into small pieces with fragment buffer at appropriate temperature. Then First-strand cDNA was generated using random hexamer-primed reverse transcription, followed by a second-strand cDNA synthesis. The cDNA fragments obtained from previous step were amplified by PCR, the double stranded PCR products from previous step were heated denatured and circularized by the splint oligo sequence to get the final library.