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How far advance can you make PCR mix? - I'm a first time poster. (Oct/17/2008 )

Hi, I'm working on RT-PCR, so I need to make RT mix, do RT, then make PCR mix, do PCR.
To save time to make 2 mixes 1.5 hours apart, I'd like to see if I can make RT mix and PCR mix at the same time.

If I make PCR mix and store it at 4 degree until RT is done (takes 1.5 hours), will it effect the result?

Very appreciate your input.

-adarc-

should be no problem (especially if you leave out the primers). just make sure you keep track of which premix is which.

-mdfenko-

i would advice you to not add in the polymerase/reverse-transcriptase enzyme until when u are really going to do the experiment.

i have a thing with enzyme . if they are not being used they should be in their glycerol solution and not chilling around in 4C. loss of activity and unwanted reaction might occur in the mix at 4 C who knows.

-Hanming86-

You can buy ready-made PCR-master mixes (without primers and template DNA), e.g. from promega. Therefore it should be no problem.

-hobglobin-