Taq polymerase can only synthesize DNA, so how do …
SOURCE: © 2006 Sumanas, Inc. KEYWORDS: Polymerase chain reaction, DNA amplification, Taq polymerase, genomics
ShineGene-Hotstart Taq DNA Polymerase-Hotstart Taq …
Because the values generated from blue/whitemethods vary significantly between individualreplicates and rely on a series of calculatedextrapolations, we have chosen to conservativelyrepresent the fidelity of Q5 High-Fidelity DNAPolymerase as >100X Taq, and Phusion HighFidelity DNA Polymerase as >50X Taq. With ever-decreasing costs and extremely large datasets,next generation sequencing techniques may soonbe able to provide direct, cost effective methodsfor more accurately quantitating error rates for anultra high-fidelity polymerase like Q5.
The discovery and development of high-fidelity polymerases has for many years been a key focus at New England Biolabs (NEB). Highfidelity amplification is essential for experiments whose outcome depends upon the correct DNA sequence (e.g., cloning, SNP analysis,NGS applications). Whereas traditional fidelity assays are sufficient for Taq and other moderately faithful enzymes, Q5, an ultra highfidelity enzyme, pushes the limits of current methods used to assess this critical feature of DNA polymerases.
The Role of Taq Polymerase in PCR | Sciencing
Taq DNA Polymerase is a thermostable DNA polymerase. It is suitable for applications requiring high temperature synthesis of DNA. Taq DNA Polymerase catalyzes the polymerization of nucleotides into duplex DNA in the 5' to 3' direction with the presence of Mg2+ and has the 5' to 3'exonuclease activity.
In this study, Q5 was examined to determine itsfidelity compared to Taq DNA polymerase usingthe two methods described below (Figure 2). A3,874 bp target was PCR amplified with eitherTaq (Thermopol Buffer), Q5 (Q5 Reaction Bufferwith or without GC enhancer) or Phusion®(Phusion HF Buffer) DNA Polymerase. Observed mutation rates were determined using both the blue/white selection method after 16 PCR cycles (4)and by Sanger sequencing after 25 PCR cycles(Table 1). The error rate per base incorporatedwas determined after calculating the effectivenumber of amplification cycles for each experiment as described previously (4, 5). Comparingthe data sets from Taq indicates that the twomethods generate similar results with error ratesof ~1 in 3,500 bases. Q5, on the other hand,yielded a significantly lower number of errorsthan Taq in both assay systems, consistent withan error rate of ~10-6. The side-by-side evaluation of Taq and Q5 using the blue/white methodsuggests that Q5 is ~200X more faithful at replicating DNA than Taq. Similar results were observed for Q5 when the GC enhancer was addedto the reactions (data not shown). For Phusion,the error rate was determined to be 80±39 timesbetter than Taq using the blue/white method and84 times Taq using the sequencing method.
The Role of Taq Polymerase in PCR
We demonstrate that despite lacking a 3'----5' proofreading exonuclease, the Thermus aquaticus (Taq) DNA polymerase can catalyze highly accurate DNA synthesis in vitro. Under defined reaction conditions, the error rate per nucleotide polymerized at 70 degrees C can be as low as 10(-5) for base substitution errors and 10(-6) for frameshift errors. The frequency of mutations produced during a single round of DNA synthesis of the lac Z alpha gene by Taq polymerase responds to changes in dNTP concentration, pH, and the concentration of MgCl2 relative to the total concentration of deoxynucleotide triphosphates present in the reaction. Both base substitution and frameshift error rates of less than 1/100,000 were observed at pH 5-6 (70 degrees C) or when MgCl2 and deoxynucleotide triphosphates were present at equimolar concentrations. These high fidelity reaction conditions for DNA synthesis by the Taq polymerase may be useful for specialized uses of DNA amplified by the polymerase chain reaction.
PCRBIO HS Taq DNA Polymerase uses advanced hot-start technology for superior sensitivity. Whether you need a hot-start assay for high throughput, automated reaction set up or for detection of a low copy number template, PCR Biosystems offers you a robust industry-leading enzyme to meet your needs.
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Taq DNA Polymerase from Bioneer
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Taq polymerase can only synthesize DNA, so how do we …
Re: How does Taq Polymerase know when to stop DNA synthesis
Taq DNA Polymerase (Thermus aquaticus)
Description Taq DNA Polymerase is a thermostable DNA polymerase
PCR Taq, Taq DNA Polymerase - GenScript
PCRBIO HS DNA Polymerase uses the latest developments in polymerase technology and buffer chemistry to enhance PCR speed, yield and specificity. The enzyme and buffer system allow for superior PCR performance on complex templates such has mammalian genomic DNA. PCRBIO HS Taq DNA Polymerase can perform consistently well on a broad range of templates (including both GC and AT rich). Due to enhanced efficiency and specificity the enzyme is perfectly suited to multiplex PCR.
Taq, Tfl, and Tth DNA Polymerases* are recombinant ..
For added convenience PCRBIO HS Taq DNA Polymerase is also available as a 2x ready mix. PCRBIO HS Taq Mix Red is suitable for direct loading on agarose gels, it contains a red dye for tracking during agarose gel electrophoresis.
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