PCR reagent autoclave - (Mar/15/2010 )

Hi,

How to autoclave PCR reagents? like MgCl2? water?

While I think it depends on the volume of medium autoclaved. But what is the boundary for the time? If the bottle or flask contains only 1/3 volume medium, we can just autoclave 15min, 120 degrees, 15psi. is that right? How about 1/2 volume medium?

Dear Rachel2010

Do not autoclave the MgCl2. Whenever we autoclave, the water amount will be either increase or decrease, and thus changes the MgCl2 concentration, and this will lead to inconsistent result in your PCR later.

If is water, you can safely autoclave. But I do prefer to use commercially purchased nuclease free water Eg from Promega.

Hope this helps.

Adrian

Why would you want to autoclave PCR reagent? You won't erradicate DNA that may be present in your solution.