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Reverse transcription of a circular RNA templateModerator: BioTeam
8 posts • Page 1 of 1
Reverse transcription of a circular RNA templateAs above, will the cDNA product be linear or circular (say if I use Superscript II or III)?
I suppose the cDNA product to be single-stranded. Am I right?
The cDNA will be circular only if you use ligase and it will be dsDNA, if you use primers for both strands of course. However, if the primers will be far enough from each other, you will get broken cirkular dsDNA
http://www.biolib.cz/en/main/
Cis or trans? That's what matters.
Re: Reverse transcription of a circular RNA templateThank you for your reply.
I use only one primer for the reverse transcription and there is no ligase added, so the cDNA should be single-stranded and linear, right? If I want to get dsDNA, can I use DNA Polymerase I to extend the other DNA chain, using the hairpin loop at the end of the first-strand cDNA as the primer? I'm a little confused about your 2nd sentence though. Are you talking about a RT-PCR?
Re:
No, I'm not talking about RT-PCR, just RT instead. The hairpin theory came from this link 'http://dwb4.unl.edu/Chem/CHEM869N/CHEM869NLinks/www.dur.ac.uk/~dbl0www/Staff/Croy/cDNAfigs.htm'. The 2nd paragraph and fig. Thank you very much.
Re: Reverse transcription of a circular RNA templateActually, I want to get ds cDNA from RNA. I've figured it out. There are several kits available that can synthesize ds cDNA from RNA (there is a step for second-strand cDNA synthesis after the first-strand synthesis). Thank you for your help.
8 posts • Page 1 of 1
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