I am trying to amplify ssDNA product using PCR. I am just wondering whether following the normal PCR protocol will yield a product since the starting DNA is not dsDNA. Have anyone tried amplifying a ssDNA or know a protocol that I should follow?
All pcr starts with ssDNA after the first denaturation step so I would either start a normal pcr without denaturing the first cycle or just treat it exactly like a normal pcr. Not denaturing might be preferable if your template is very long but short ss dna is more robust and just treat it as a normal pcr
Yes, it will work fine. This is what happens during all reverse transcription PCRs as the initial template (first strand cDNA) is single stranded. I would not skip the denaturation step because depending on the template, the ssDNA may form a secondary structure, which the denaturation step would melt.
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