whats a good, typical final primer conc?

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S.A.D.
S.A.D.'s picture
whats a good, typical final primer conc?

Hi,

I'm just curious, does anyone know for PCR in general, what would be a good, safe and typical final concentration of primers in the PCR reaction tube.

Also what concentration range would be too little, and too much.

I'm also wondering what happens in general if I have an excess of primers in the reaction?

Thanks

SAD

R Bishop
R Bishop's picture
I make up my stock primers at

I make up my stock primers at 10uM and use 1 ul per 10ul PCR reaction, so a final concentration of 1uM works fine. In cases for high background bands you can go as low as 2uM stock concentration and still use a 1:10 dilution for a final concentration of 0.2uM. Im sure people use even less than that with success.

Rb

Fraser Moss
Fraser Moss's picture
R Bishop wrote:I make up my

R Bishop wrote:

I make up my stock primers at 10uM and use 1 ul per 10ul PCR reaction, so a final concentration of 1uM works fine. In cases for high background bands you can go as low as 2uM stock concentration and still use a 1:10 dilution for a final concentration of 0.2uM. Im sure people use even less than that with success.

Rb

I agree

I generally use 1ul of 10 pmol/ul (10uM) stock in a 50ul PCR = 0.2uM final conc of each primer in the PCR reaction.