what is the annealing and elongating temperature of a specific primer (31 bases) for reverse transcription ?
Dear Mohamed!
First of all, you should look at the manufacture's protocol, because conditions may vary from one manufacture to another one. It may also depend on plans for further work with cDNA. You should also estimate GC-content in the target transcript. Many manufacturers do not recommend annealing a specific primer separately, but sometimes it is the important step. Prepare the reaction mixture, add your primer and RNA. The temperature of typical reaction is 50-55 °C (for most kits that I used with specific primers). For transcription of GC-rich RNA, the reaction temperature can be increased to 65 °C . But I think ,that it will work in the ordinary conditions.
Hi,
I hope this helps;
https://www.sigmaaldrich.com/technical-documents/articles/biology/reverse-transcription.html
you should simply follow the protocol of your prefered enzyme, usually they are really good and do explain which temps should be used regardless of primer composition. I prefer Superscript 4.
https://assets.thermofisher.com/TFS-Assets/LSG/manuals/SSIV_Reverse_Transcriptase_UG.pdf
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