PCR primer design, real time versus old school - (Oct/30/2007 )

Hello!

I have what is probably a very stupid question. I need to design primers for regular RT-PCR (not real time), I have found published sequences for all my targets but they were all used in real time. Will this also be ok for regular old school PCR? My understanding is that yes it will but doing it the other way round may be an issue as in real time interactions with the probe can be an issue but I thought I should check before I either spend loads of time designing my own or order ones from the published sequences and them not work!

Cheers!

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Hi, as far as i know real time primers should definitely work in standard "old school" PCR, the only thing i would suggest is to check the product size, usually amplicons are smaller for real time qPCR, which might make it more difficult to detect by standard agarose, gel, but as long as they are over 100bp you should be fine.
i'd say go ahead and get the published ones unless you are looking into something different.
oh! you might need to change the amplification parameters, but whatever they used for real time will probably work too.

Good luck!

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Thanks for you help, I shall check the sizes as you suggested and give it a go.

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