( /subof PfuDNA polymerase are not un...)Pfu DNA Polymerase as a Hot Start Alternative

/sub>of Pfu DNA polymerase are not unusual when one considers the natural habitat from which it was derived. Pfu DNA polymerase was cloned from the hyperthermophilic archaeon Pyrococcus furiosus, which grows optimally at 100C in geothermal marine sediments.⁴ In contrast, Taq DNA polymerase was cloned from the thermophilic eubacterium Thermus aquaticus, which grows optimally at 70C in thermal springs.⁴

Activity vs. Temperature Profiles

Figure 1

In order to demonstrate the difference in activity vs. temperature profiles between Taq and Pfu DNA polymerases, the polymerase activity of the two enzymes was compared over a range of temperatures encountered in PCR amplifications. The activities of cloned Taq and cloned Pfu DNA polymerases were compared at equal unit concentrations at temperatures ranging from 15 to 100C. The percent of maximal polymerase activity is plotted as a function of temperature in figure 1. Taq DNA polymerase exhibited optimal (84%) polymerase activity between 60 and 74C and 27% to 70% activity between 40 and 50C. Pfu DNA polymerase, on the other hand, exhibited optimal activity between 70 and 77C and only 2% to 8% activity between 40 and 50C. No significant differences in activity vs. temperature profiles were observed between cloned and native DNA polymerases (data not shown). Polymerization by Pfu DNA polymerase decreased dramatically at temperatures above 75 to 80C, presumably because the primer-template duplex dissociates at elevated temperatures. The T_m of the 40-base primer used in the primer-extension assay was calculated to be approximately 92.6C.⁵

'"/>

Source:

Page: All 1 2 3 4 5 6 7 8 9
Related biology technology :

1. Novel PCR Enhancing Factor Improves Performance of Pfu DNA Polymerase
2. PCR Performance Comparisons Between pfuturbo and Taq DNA Polymerases
3. pfuturbo DNA Polymerase: A High-Performance, High-Fidelity Enzyme Ideal for PCR Cloning
4. Unique DNA Polymerase Formulation Excels in a Broad Range of PCR Applications
5. Enhanced Amplification of Long Targets with PfuTurbo DNA Polymerase
6. High-Fidelity PCR with a Novel Polymerase Mixture
7. Optimizing pfuturbo DNA Polymerase Amplification Reactions with Perfect Match PCR Enhancer
8. Unique Enhanced DNA Polymerase Delivers High Fidelity and Great PCR Performance
9. The Use of MasterTaq Polymerase for PCR with Humic Material-Contaminated DNA
10. HotStarTaq DNA Polymerase
11. C. therm. Polymerase One-Step RT-PCR System