Additional file 1.
Transcription of the PolB gene of HcDNAV. To verify the transcription of the HcDNAV PolB gene, reverse transcription-PCR (RT-PCR) experiment was conducted. The total RNA samples were isolated from HcDNAV-inoculated Heterocapsa circularisquama cells collected at 0, 1, 6, 12, and 24 hpi, then reverse-transcribed according to the method given by Nagasaki et al. . PCR amplification was performed using a DNA polymerase KOD FX (Toyobo, Osaka, Japan) and primers designed for amplification of HcDNAV PolB gene fragment (01F: ACG TTT TAA ATG ATG TTA TTA ATG, 01R: GCC ATT TTA ATA TAT GAA TAA A); the reaction cycling conditions were 94°C for 2 min, then 25 cycles of 98°C for 10 s, 50°C for 30 s, 68°C for 1 min. Lanes 1 to 5 show the RT-PCR fragments amplified from cDNAs at 0, 1, 6, 12, and 24 hpi, respectively, and lane 6 shows the PCR fragments amplified from HcDNAV DNA (positive control).
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Ogata et al. Virology Journal 2009 6:178 doi:10.1186/1743-422X-6-178