What precautions should be taken when using inosine-containing primers?

TaKaRa Taq DNA Polymerase and TaKaRa Taq DNA Polymerase Hot Start Version are compatible with inosine-containing primers. Inosine-containing primers should not be used with PCR enzymes that have 3'-5' exonuclease activity (e.g., PrimeSTAR HS DNA Polymerase, PrimeSTAR Max DNA Polymerase, PrimeSTAR GXL DNA Polymerase, TaKaRa Ex Taq DNA Polymerase, or Takara LA Taq DNA polymerases), nor with Terra [...]

How should oligos be purified for PCR?

Standard desalted primers are satisfactory for most PCR applications.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 [...]

How do I calculate the melting temperature (Tm) of primers?

The primer melting temperature (Tm) is the estimate of DNA-DNA hybrid stability. Knowing the Tm is critical for determining an appropriate annealing temperature (Ta). A Ta that is too high will result in insufficient primer-template hybridization, leading to low PCR product yield. A Ta that is too low may lead to non-specific product amplification. Calculation of the Tm of primers shorter than [...]

What is the storage temperature of Premix Ex Taq™ (Probe qPCR)?

Premix Ex Taq™ (Probe qPCR) is protected from light and stored at -20°C for long term storage. Once thawed, it should be stored at 4°C for up to 6 months." ProdStorageReal-time PCR [...]