Should I use Oligo-dT with a high processivity reverse transcriptase to make the total mRNA and then use specific primers to PCR amplify it? or is there a better way?
Thanks
Hi Ela
As you mentioned, using OligodT for cDNA conversion is the most commonly used method worldwide and very rarely researchers will use gene specific reverse primer for RT reaction. This method is wildly used while walking, RACE protocols for avoiding non specific amplifications in the subsequent down stream steps. But in your case, I feel OligodT RT reaction will be the most appropriate one because you need to amplify full length of the gene. If you go with GS reverse primer for RT, you may end up in partial length 3' region in your full length gene and if your 3' UTR region is very important, then again you need to go for RACE protocol.
Good luck
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