Hi everyone,
We are planning a RNA analysis of expression of mRNA of an interest protein. The primers that appear to be the most reliable are Taqman primers, suggested to work better with Taqman Polymerase Mastermix.
However, we already have a master mix from Agilent (PfuUltra II HS DNA polymerase). Has anyone tried using the Taqman primers with other polymerase types with success? We are afraid to ask the manufacturers as, of course, they will prefer us to buy their own polymerase.
Thank you in advance.
Primers are primers whoever designs/manufactures them therefore will be compatible with any polymerase. Taqman primers will have been chosen for their robustness (specificity and efficiency) from a number of primer/probe sets so whilst there may be different [Mg2+] [other PCR enhancers] in each of the master mixes I would wager that the PCR conditions with each would be exactly the same!
You can choose a wide range of compatible polymerases with hotstart feature in-built. However, the most important point to note is the content of the master mix especially Mg2+, Na+ and other molecules being used as PCR enhancers. Else, you can do a Mg2+ titration using the same master mix, your new polymerase, taqman primers and your positive template to achieve the desired RFU.
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