Detection of positive and negative KRBV genomic RNA strands as an indication of viral replication. Detection of positive and negative KRBV genomic RNA.

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Detection of positive and negative KRBV genomic RNA strands as an indication of viral replication. Detection of positive and negative KRBV genomic RNA strands as an indication of viral replication. (A) Amplification of KRBV sequence with and without RT with KRBV-specific primers. The first lane has KRBV RNA subjected to RT-PCR, the second lane has KRBV RNA subjected to PCR only, the third lane has KRBV cDNA subjected to PCR, the fourth lane shows the unamplified input cDNA, and the last lane shows the nontemplate control subjected to RT-PCR. (B) RT-PCR of DNase-treated KRBV RNA. T, DNase treated; NT, not treated. (C) RT-PCR amplification of Anopheles ISF sequences with forward only (F), reverse only (R), or both virus-specific primers during RT. Top gel, KRBV; bottom gel, from left to right, DSwV, McPV, and HaCV. (D) PCRs of cDNA generated with forward primer KRBV41F and with downstream (KRBV42R and KRBV3UTR1F/1R pair) and upstream (KRBV20F/R pair) primers and control RT-PCR of KRBV RNA for upstream primers. Agathe M. G. Colmant et al. mSphere 2017; doi:10.1128/mSphere.00262-17