What temperature is considered optimal for Taq polymerase activity
Taq' s optimum temperature for activity is 75–80°C, with a half-life of greater than 2 hours at 92.5°C, 40 minutes at 95°C and 9 minutes at 97.5°C, and can replicate a 1000 base pair strand of DNA in less than 10 seconds at 72°C.ne of Taq' s drawbacks is its lack of 3' to 5' exonuclease proofreading activity resulting in relatively low replication fidelity. Therefore it replaced the DNA polymerase from E. coli originally used in PCR [7]. Taq's temperature optimum for activity is 75-80 0 C, with a half-life of 9 minutes at 97.5 0 C, and can replicate a 10 3 base pair strand of DNA in less than 10 seconds at 72 0 C [8].The incorporation of dNTPs into DNA is catalyzed by Taq DNA polymerase I.aq DNA polymerase has among the highest error rates of the thermophilic polymerases at 285 x 10 -6 errors per template nucleotide. Tli polymerase has a proof reading ability that is five-fold better than Taq at 57 x 10 -6 errors per template nucleotide and Pfu polymerase also demonstrates fidelity in this range [10]. The most commonly used DNA polymerase for PCR is Taq DNA polymerase, which has an error rate of approximately 1 × 10 –5 errors per base. This error rate is relatively high due to the enzyme's lack of 3′→5′ exonuclease (proofreading) activity.aq DNA polymerase is suitable for most PCR applications that do not require a high-fidelity enzyme, such as detecting specific DNA or RNA sequences. The error rate of Taq DNA polymerase is approximately 1 × 10 –5 errors/base, although the fidelity does depend somewhat on the reaction conditions. One of Taq' s drawbacks is its lack of 3' to 5' exonuclease proofreading activity resulting in relatively low replication fidelity.Originally its error rate was measured at about 1 in 9,000 nucleotides.ne of Taq' s drawbacks is its lack of 3' to 5' exonuclease proofreading activity resulting in relatively low replication fidelity. Size and activity: While variable weights have been reported, the approximate size of Taq is 94 kd, with the activity of a DNA polymerase localized to the C-terminus and 5’ to 3’ exonuclease activity localized to the N-terminus.aq requires the presence of all four species of deoxyribonucleoside triphosphates and DNA for optimum catalytic activity. pH dependency: The pH optimum for the enzyme is within the range of 7-8 pH units when at 80 °C, and will vary depending on the buffer system used. T. aquaticus is a bacterium that lives in hot springs and hydrothermal vents, and Taq polymerase was identified as an enzyme able to withstand the protein-denaturing conditions (high temperature) required during PCR.Therefore it replaced the DNA polymerase from E. coli originally used in PCR.ne of Taq' s drawbacks is its lack of 3' to 5' exonuclease proofreading activity resulting in relatively low replication fidelity. Characterization of Thermus aquaticus DNA Polymerase Functionality: Utilizing the inherent 5’ to 3’ exonuclease activity of Taq, researchers are able to simultaneously achieve PCR amplification and signal release from a target-specific fluorogenic probe.aq requires the presence of all four species of deoxyribonucleoside triphosphates and DNA for optimum catalytic activity. pH dependency: The pH optimum for the enzyme is within the range of 7-8 pH units when at 80 °C, and will vary depending on the buffer system used. In the presence of Mn 2+ , Tfl DNA polymerase can use RNA as a template. Tfl DNA polymerase exhibits a 5′→3′ exonuclease activity but lacks a 3′→5′ exonuclease activity. This enzyme is commonly used in PCR (Gaensslen et al. 1992), where its activity is similar to that of Taq DNA polymerase.Tfl DNA polymerase is used in the Access and AccessQuick™ RT-PCR Systems.aq DNA polymerase is suitable for most PCR applications that do not require a high-fidelity enzyme, such as detecting specific DNA or RNA sequences. The error rate of Taq DNA polymerase is approximately 1 × 10 –5 errors/base, although the fidelity does depend somewhat on the reaction conditions. Temperature dependency: Thermophiles are prevalent in nature and certain prokaryotic species thrive at temperatures above 45 °C. The temperature-dependency of Taq makes it optimum at 80 °C, where its catalytic activity is more than ten times that typically observed at 37 °C.It is possible that the decrease in activity above 80 °C is actually due to the denaturation of double-stranded DNA at these temperatures.aq requires the presence of all four species of deoxyribonucleoside triphosphates and DNA for optimum catalytic activity. pH dependency: The pH optimum for the enzyme is within the range of 7-8 pH units when at 80 °C, and will vary depending on the buffer system used. Stoffel Fragment differs from AmpliTaq® DNA Polymerase in that it is more thermostable (by approximately 2-fold), exhibits optimal activity over a broader range of magnesium ion concentration (2 mM-10 mM) and lacks intrinsic 5' to 3' exonuclease activity.n the DNA Thermal Cycler 480 and the DNA Thermal Cycler, the average block temperature is used to time the incubation periods. At least 1 minute must be specified for sample temperature equilibration for the most reliable amplification in standard 0.5 mL GeneAmp® PCR Reaction Tubes.
optimal temperature for activity is 75–80°C, with a half-life of greater than 2 hours at 92.5°C, 40 minutes at 95°C and 9 minutes at 97.5°C.