standard PCR efficiency - No real time! (Apr/07/2008 )

hello everyone.
Does anyone have a clue as to how to measure the efficiency of standard PCR (i.e. PCR followed by electrophoresis). Any suggestions will be more than appreciated.

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Never done, but I would guess can be done by measuring the yield after the PCR and relate it to number of cycles. And yield via DNA concentration (?) before after PCR...
Have also a look here:
(Calculations for Molecular Biology and Biotechnology p 147 ff.)

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You can do it by amplifying serial dilutions of your target and after running the electrophoresis measuring the intensity of the bands and plotting it together (target concentration vs band intensity). Just be sure that you're stopping the reaction when it's still in the linear phase of amplification, for that you need to find out the number of cycles at which the reaction gets saturated....

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