I got a custom made sequence detection primers with 10 nm scale and a probe with 6 nm scale for a microbial sequence and I did not get anything else from the company. I want to know how to set the reaction if I am using a universal qPCR master mix and the template is bacterial DNA. What is the final concentration used for the probe and primers. Should it be as in conventional PCR? 0.5 uM or what?
Wolfgang Schechinger
Yes, just start with the usual concentrations as in any PCR.
The company should have written somewhere how much there is actually in the vial. Usually, it's either in the synthesis report or simply printed on the sticker, so you may calculate the amount of water that you have to add to dissolve the dry material.
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