RT-PCR technique

alex dobrovic adobrovic at medicine.adelaide.edu.au
Tue Mar 3 19:56:42 EST 1998

One big advantage of a 2 step technique is that multiple PCRs can be used
to amplify multiple RNAs,

>Hey again,
>Part two! I have used the Promega Access RT-PCR kit, with minor
>modifications, for my RT-PCR experiments. I have done several reactions
>mixing and matching buffers, RTs and differnt DNA polymerases (Tfl, Taq,
>Tth) to come up with the best combination.
>Are there any inherent advantages to a two-step RT/PCR method (e.g.
>SuperScript RT followed by Taq PCR) as opposed to a one-step RT-PCR
>method (e.g. AMV-RT/Tfl combination)? Which would you recommend? I am
>not particularly concerned with generating full-length transcripts,
>hence I use AMV-RT and gene-specific primers as opposed to SuperScript
>and an oligo(dT)n primer.
>Any help/advice/support is much appreciated
>Jonathan Shillingford



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