What is my control when running a PCR reaction? - (Oct/12/2008 )

Hi I am running a PCR reaction and wondered what control if any I should use. It is part of my degree at university so we have to do it all thoroughly. We have been asked to find out what the optimum Mg concentration is so should I run a reaction without adding any Mg and use that as my control or is it something to do with another part of the reaction... primers etc?

Thanks in advance and hope it makes sense.

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Always include a NTC = no template control

usually you then do a mg-titration (but omit the 0mM mgcl2, the pcr won`t run under these conditions.

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It also would be good to include a positive control to evaluate if the PCR itself was successful. Sometimes when a PCR produces no product with your samples doesn't necessarily mean that the PCR run didn't work....but if the positive control doesn't have a product than you know that something was wrong with your reaction.

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