Hi!
Has anyone worked with negative-sense RNA viruses? I need to express recombinant proteins using a vector, so it will be necessary to start with ssRNA(-) to obtain cDNA for selecting the gene of interest. Are there kits with RNA-dependent RNA polymerase to obtain the positive-sense strand (necessary for the following step with the reverse transcriptase), or do I need to isolate the enzyme in some way? Thank you.
When making cDNA the sense of the strand does no matter as you use random hexamers to prime. Companies like ThermoFisher and NEB sell both the random hexamers and reverse transcriptase. Once you have the cDNA you can then use regular PCR to generate the other strand.
For reverse transcriptase is not important the type of matrix (sense or not sense). You can produce with random hexamer you copy DNA and then with PCR to receive PCR product for cloning.
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