dNTP Mix

Bio-Rad dNTP mix remains stable after precycling. dNTP mix was precycled for 25 rounds of thermal cycling, then incubated with additional reaction components for 25 cycles of PCR. Genomic DNA template (20 ng) was amplified with iTaq™ DNA polymerase using primers to the HFE gene. Two products, one 500 bp and the other 2 kb in length, were both produced in similar amounts regardless of whether the dNTPs were precycled (lanes 2 and 4) or not (lanes 1 and 3). Lane M, 500 bp EZ Load™ molecular ruler.

Bio-Rad dNTP mix performance in real-time PCR remains unchanged after precycling. dNTP mix was precycled for 25 rounds of thermal cycling, then incubated with additional reaction components for 25 cycles of real-time PCR. Tenfold serial dilutions of 500 ng to 50 pg of genomic DNA were amplified using primers to the IL-1β gene. At each concentration, reactions were performed in triplicate. Amplification was performed with iTaq™ DNA polymerase. Amplification was successful whether the dNTPs were precycled (■) or not (■).

Bio-Rad dNTP mix outperforms an alternate supplier's dNTP mixture in real-time PCR. Tenfold serial dilutions of 500 ng to 50 pg of genomic DNA were amplified using primers to the IL-1β gene. At each concentration, reactions were performed in triplicate. Amplification was performed with iTaq DNA polymerase and either the Bio-Rad dNTP mix (■) or the alternate supplier's dNTP solution (■). The Bio-Rad dNTP mix is a more robust mixture, producing more consistent C_T values than the other dNTP solution at low input DNA concentrations.