What primer concentration should be used for PCR?
What primer concentration should be used for PCR?
The final concentration of each primer should be between 0.1 and 0.5 µM. A stock solution of each primer is typically 10–20 µM.
-For PCR amplicons less than 10 kb, 0.2 µM produces satisfactory results.
-For amplification of long targets (~17 kb) with Takara LA Taq DNA polymerases or Takara Ex Taq DNA polymerases, the primer concentration can be increased up to 1 µM.
-The recommended primer concentration for high-yield polymerases such as Advantage 2 DNA polymerases and Titanium Taq DNA polymerases is 0.4 µM.
Primer concentrations that are too high increase the chance of mispriming, which may result in nonspecific amplification. Primer concentrations that are limiting can result in extremely inefficient amplification.