Transcriptomic Analysis of Calonectria pseudoreteaudii
Briefly, mRNA of Calonectria pseudoreteaudii was enriched using the oligo(dT) magnetic beads. Adding the fragmentation buffer, the mRNA was interrupted to short fragments (about 300 bp), then the first strand cDNA was synthesized by random hexamer-primer using the mRNA fragments as templates. Buffer, dNTPs, RNase H and DNA polymerase I were added to sythsize the second strand. The double strand cDNA was purified with QiaQuick PCR extraction kit and washed with EB buffer for end repair and single nucleotide A (adenine) addition. Finally, sequencing adaptors were ligated to the fragments. The required fragments was purified by agrose gel electrophoresis and enriched by PCR amplification.