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. Author manuscript; available in PMC: 2020 Oct 1.
Published in final edited form as: J Virol Methods. 2019 Jul 14;272:113701. doi: 10.1016/j.jviromet.2019.113701

Figure 3. Primer-independent cDNA synthesis of viral RNA impairs standard RT-PCR strand-specificity.

Figure 3.

100 ng of in-vitro synthesized RNAs corresponding to the genomic (G) and antigenomic (AG) segments of L, M, and S RNAs and intracellular RNAs from RVFV-infected cells (MP-12 IC) were used for cDNA synthesis without the use of specific RT primers. (A) After cDNA synthesis, the samples underwent PCR using unmodified PCR primer sets, specific for genomic L (lanes 2-4), M (lanes 5-7), and S (lanes 8-10) segments. (B) Experiments were done similar to (A), except that the samples underwent PCR using unmodified PCR primer sets, specific for antigenomic L (lanes 2-4), M (lanes 5-7), and S (lanes 8-10) segments. Lane 1 in (A) and (B) represents the DNA size marker, and the location of the markers having 100 and 200 base pairs (bp) bands are indicated by arrows. The PCR products were analyzed by agarose gel electrophoresis.