Optimized RT–PCR protocol applied to TDV-2. (A) RT–PCR of TDV-2 RNA using optimized RT and PCR annealing temperatures. (B) Bioanalyzer electropherogram plot of scaled up PCR reactions. (C) Bioanalyzer electropherogram plot of TDV-2 PacBio library. The 50 and 17 000 bp peaks in the electropherogram plots are the kit markers. Abbreviation: FU, fluorescent unit.