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I am trying to design primers for a single stranded DNA virus. But I am having a dilemma as whether we design the primers same way as double stranded DNA?
do we have to design forward and reverse primers both or just a single primer forward or reverse works?

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If you've e.g. a parvovirus you design primers as normal, i.e. must be a pair of primers as for double stranded DNA. Albeit there's only one strand present after the first round of PCR there both strands will be present and without the reverse primer the PCR can't work. If you want evidence of this, you've had a SARS-CoV-2 diagnosis? Well thats a forward and reverse primer PCR starting with ssRNA converted to cDNA, i.e. single copy and then a pair of primers PCR the resulting locus.

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    $\begingroup$ Even long before covid, starting from single-stranded DNA is a standard PCR technique, for example gene cloning from extracted cDNA. The principle of the chain reaction does not change, you always need two primers for amplification. Else, you always amplify from the original template over and over again, but this is not exponential and you'd produce an excess of forward (or reverse) strand product which has no use. $\endgroup$
    – ATpoint
    Sep 20, 2023 at 14:08
  • $\begingroup$ So we will just remove both the denaturation steps from PCR conditions, and the rest of the steps remains same, is it right? $\endgroup$
    – Maharshi
    Sep 21, 2023 at 6:32
  • $\begingroup$ No, everything is the same. Exact same. Get local supervision for lab work, this is a bioinformatics community. $\endgroup$
    – ATpoint
    Sep 21, 2023 at 6:47

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