Transcription of PCR products without a primer coupled with promotor sequence - (Dec/02/2012 )
Is there a way to transcribe PCR products wihtout using a primer that has a transcription promotor. I plan to do base specific cleavage followed by MALDI TOF for strain differentiation. My PCR is a multiplex PCR with 15 primer pairs. Thus, tagging the primers is not a possibility.
Any alternatives are most welcome.
Why do you need to transcribe? surely this is essentially the same as doing a PCR with Uracil.
All protocols for MALDI TOF based sequencing using base soecific cleavage is by this method. The cleavage is by RNAses, I think that is the reason.
Ah, fair enough. You would definitely need a promoter to get this to work. Would it be possible to clone (possibly topo-TA) the PCR products?
That is possible. However, if there are quicker methods, it is better as the final objective would be to see if the method can be diagnostically applicable. As cloning would take additional days is not the best option.