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Primer Dilution. ...water or buffer? - (Jun/01/2007 )

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QUOTE (Kupac @ Jul 26 2007, 05:46 AM)
I've used both water and TE. At first I was concerned that EDTA would chelate Mg++, and reduce the activity of Taq, but actually it works fine. Probably the EDTA concentration is not high enough for that, but it still protects my primers. So I just use TE from now on.

Usually I dilute my primer with TE buffer. And it works just as fine. I guess you are right, the EDTA concentration is low. Plus if you want to be sure, you can try to use TE with reduced EDTA (available commercially). laugh.gif


I always prefer water to be on the safe side because you can never know the exact effects of extra ions or EDTA on your PCR. If you dilute some amount of your primers and aliquote them, you can store the stocks in -80 and thaw again only when it's necessary. This will also protect your primer stockts from degradation and/or contamination.


We generally dilute our primers in a very low concentration of TE buffer, to keep the pH consistent.


It is fine.

But I make my primers stocks at two different dilution.
The main freezer stock at 100uM is made using TE and the working Primer stocks, 10uM (that actual goes into the PCR reaction) is made by diluting the 100uM freezer stock with ds water. (MiliQ)


QUOTE (Doyo @ Jun 2 2007, 12:48 AM)
Hi guys...

I am currently using 10X PCR buffer containg MgCl2...I did not used this buffer to dilute my primers but water. To avoid unecessory calculation (for Mg and others) in preparing master mix. My friend is not happy on this...of course my primers are working wel.

What do you advice me?


I use nuclease free water


we use steriled H2O milliq to dilute primer.never I had problem whit that. Best Regard.


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