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Could have more DNA concentration inhibition effect on PCR? - (Sep/20/2008 )

Hello Dears,

I got a new assay (SNP-Stream) which needs 2ng DNA for 48-plex Multi-PCR (5 uL Total volume). My colleges used allways 6 ng DNA. Now with this protocol I got bad results. Could the more DNA concentration have an inhibition effect on PCR?! The expriment is too expensive that I can not test it with different DNA concentrations in our lab :-(
My be you can answer me before I am forced to do it.

Thanks a lot
DNAvector

-DNAvector-

QUOTE (DNAvector @ Sep 20 2008, 03:37 PM)
Hello Dears,

I got a new assay (SNP-Stream) which needs 2ng DNA for 48-plex Multi-PCR (5 uL Total volume). My colleges used allways 6 ng DNA. Now with this protocol I got bad results. Could the more DNA concentration have an inhibition effect on PCR?! The expriment is too expensive that I can not test it with different DNA concentrations in our lab :-(
My be you can answer me before I am forced to do it.

Thanks a lot
DNAvector


Too much DNA can indeed inhibit a pcr reaction (Just like too many primers). But I don't know for sure if this will happen if you use 6 ng in stead of 2 ng, but then again, the total volume is quite small (I usually use 20 ┬Ál, so then there is less impact).

-aspergillie-

Thanks aspergillie,

I think also that 6ng DNA would not be a big matter in 5uL total volume of PCR mastermix.
I dicided to testet it any way. When I got results, I would inform you as well.

Thanks a lot
DNAvector

-DNAvector-

Dear aspergillie ,

You are right. In this case (low volume and low DNA-Concentration) is no PCR Inhibition because of higher DNA amount.

Thanks
DNAvector

-DNAvector-