Invitrogen was founded in 1987 by lyle turner, joe fernandez, and william mcconnell and was incorporated in 1989. Platinum taq dna polymerase is a recombinant taq polymerase complexed with a proprietary antibody that blocks the polymerase activity at ambient temperatures. Data of expression and purification of recombinant taq dna. Taq dna polymerase is withheld until the reaction temperature is at 80c, to ensure high specificity of the products being synthesized. Add all components as in the basic pcr protocol, except for the taq dna polymerase. The hot start property of the enzyme preparation is conferred by thermolabile monoclonal antibodies that render taq dna polymerase inactive until the initial pcr denaturation step. We previously described in a paper published in bios an undergraduate lab activity. The enzyme consists of a single polypeptide with a molecular weight of 94 kda. Invitrogen platinums taq dna polymerase high fidelity. Invitrogen platinum ii hotstart pcr master mix 2x offers platinum ii taq hotstart dna polymerase premixed with platinum ii pcr buffer and dntps for convenient pcr setup.
Dna polymerases, particularly the taq polymerase used in pcr, often add an extra nucleotide to the 3. Choose exceptional sensitivity, specificity, and yield. Use platinum taq dna polymerase for the amplification of dna from complex genomic, viral, and plasmid templates, as well as in rtpcr. The enzyme is a taq dna polymerase supplied in a proprietary formulation containing 50% glycerol with buffers designed for enhanced amplification. The enzyme has terminal transferase activity which results in the addition of a single nucleotide adenosine at 3 end of the extension product. Mix contents of tube and overlay with 50 l of mineral or silicone oil. The polymerase chain reaction pcr is a powerful and sensitive technique for dna amplification 1. Taq polymerase, frequently used for standard pcr applications, exhibits 5. Efficiency comparison of seven different taq polymerases used in hemogenetics j. Platinum taq dna polymerase high fidelity contains recombinant taq dna polymerase, pyrococcus species gbd polymerase, and platinum taq antibody. Takara taq dna polymerase is a recombinantversion taq polymerase derived from the thermus aquaticus yt1 strain, and is suitable for routine pcr applications for individual reaction setup and optimization, use individual components of enzyme, dntp mixture, and 10x reaction buffer with or.
Activity is restored after the initial denaturation step in pcr cycling at 94c, providing an automatic hot start and offering increased sensitivity, specificity, and yield. Efficiency comparison of seven different taq polymerases. Taq dna polymerase is the industry standard for routine pcr. This enzyme allows amplification of simple and complex dna templates over a large range of target sizes and provides 6x higher fidelity over taq. Links to pubmed are also available for selected references. Scientists realized that thermostable heatstable dna polymerases would be needed for pcr to work efficiently. It ensures higher sensitivity, longer pcr products and higher yields compared to conventional taq dna polymerase. Taq dna polymerase pcr buffer thermo fisher scientific. Taq dna polymerase, recombinant thermo fisher scientific. Aug 20, 2016 in this article, the taq dna polymerase expression plasmid is reconstructed and the protein product is obtained by rapid purification, rapid purification of highactivity taq dna polymerase pluthero, 1993, singlestep purification of a thermostable dna polymerase expressed in escherichia coli desai and pfaffle, 1995. Taq dna polymerase is heatstable and synthesizes dna at elevated temperatures from singlestranded templates in the presence of a primer. Hotstart pcr activation approaches allow users to minimize nonspecific amplification while increasing target yield and specificity. Taq polymerase, being thermostable, proved ideal for pcr. The hot start property of the enzyme is conferred by thermolabile monoclonal antibodies that render taq dna polymerase inactive until.
Pcr enzymes and master mixes go online to view related products. Properties of thermostable dna polymerase yufei tu discoveryhistory of taq dna polymerase the original report of this enzyme, purified from the hot springs bacterium thermus aquaticus, was published in 1976. Efficiency comparison of seven different taq polymerases used. Pcr, realtime pcr, reverse transcription, and cloning taq dna polymerase and taq pcr core kit for standard and specialized pcr applications qiagen pcr buffer for minimal optimization additional readytoload pcr buffer for faster and easier handling qsolution, an innovative pcr additive, for amplification. Taq dna polymerase fast and robust pcr neb is proud to taq dna polymerase neb is proud to represent taq dna polymerase on the basis that it is an enzyme of the highest quality and value. Our jumpstart taq dna polymerase is an antibody inactivated hotstart enzyme. Themagnesium concentrationdoesnot need to be adjusted. Purification of thermostable dna polymerases gene and cell. Taq dna polymerase is appropriate for use in the amplification of dna from complex genomic, viral, and plasmid templates, rtpcr, sequencing ssdna, and cycle sequencing. Taq dna polymerase is recombinant taq dna polymerase complexed with a proprietary antibody that blocks polymerase activity at ambient temperatures. Neb provides high quality recombinant taq at an exceptional value. This enzyme has both a 5 3 dna polymerase and a 5 3 exonuclease activity but lacks a 3 5 exonuclease activity. Invitrogen platinum taq dna polymerase is a convenient and reliable hot start thermostable dna polymerase for pcr that provides enhanced specificity over that of taq dna polymerase.
Rapid purification of highactivity taq dna polymerase. For multiple reactions, prepare a master mix of components common to all. Three different dna target varying in length pah 203 bp, eng9 293 bp and eng 5 480 bp were evaluated. Activity is restored after the denaturation step in pcr cycling at 94c, thereby providing an automatic. The polymerase chain reaction pcr method for amplifying selectively discrete segments of dna has found widespread applications in molecular biology, due in part, to the substitution of a thermostable dna polymerase isolated from thermus aquaticus taq 1 for the previously used e. Dreamtaq dna polymerase uses the same reaction set. Properties of thermostable dna polymerase yufei tu discoveryhistory of taq dna polymerase the original report of this enzyme, purified from the hot springs bacterium thermus aquaticus, was. Janica b a central forensic laboratory of the police, warsaw, poland.
Taq dna polymerase is a thermostable enzyme that synthesizes dna from singlestranded templates in the presence of dntps and a primer. The polymerase chain reaction pcr was developed by chemist kary mullis in the 1980s, as a means to make many copies of dna fragments. The following guidelines are provided to ensure successful pcr using nebs taq dna polymerase. Genscript taq dna polymerases are highly thermostable recombinant dna polymerases and ideally suited for routine pcr reactions. Invitrogen platinum taq dna polymerase fisher scientific.
It is supplied with 10x standard taq reaction buffer, which is detergentfree and designed to be compatible with existing assay systems. Platinum taq dna polymerase high fidelity offers the best of both yield and fidelity and therefore, is a good choice when quantity and quality are important. Taq dna polymerase is an enzyme widely used in pcr 2. Platinum ii taq hotstart dna polymerase is designed for universal primer annealing and fast, easy pcr with its unique combina. Full text is available as a scanned copy of the original print version.
Onetaq dna polymerase is an optimized blend of taq and deep vent dna polymerases for use with routine and difficult pcr experiments. One unit of taqdna polymerase is the amount of enzyme required to incorporate 10 nmoles of deoxyribonucleotide into dna in 30 minutes at 74c. Pluthero 1993 rapid purification of highactivity taq dna polymerase. In hotstart pcr, taq polymerase is inactive until heated. Taq dna polymerase is a thermostable dna polymerase that possesses a 5. Get a printable copy pdf file of the complete article 394k, or click on a page image below to browse page by page. Even lowabundance dna templates and difficult gcrich. It has a 53 dna polymerase activity and a 53 exonuclease activity. Dreamtaq dna polymerase does not incorporate dutp, description thermo scientific dreamtaq dna polymerase is an enhanced taq dna polymerase optimized for all standard pcr applications. The enzyme consists of a single polypeptide with a molecular weight of approximately 94 kda. In this article, the taq dna polymerase expression plasmid is reconstructed and the protein product is obtained by rapid purification, rapid purification of highactivity taq dna polymerase pluthero, 1993, singlestep purification of a thermostable dna polymerase expressed in escherichia coli desai and pfaffle, 1995. Determination of taq polymerase units using an authentic taq polymerase standard. Pcr the polymerase chain reaction pcr is a powerful and sensitive technique for dna amplification 1. Part one 3 the use of the taq polymerase not only simplified the pcr procedure see above but significantly increased the specificity and the overall yield of the reaction.
The company initially found success with its kits for molecular cloningnotably, the librarian, a kit for making cdna libraries, and the fasttrack kit. Taq polymerase is a thermostable dna polymerase i named after the thermophilic eubacterial microorganism thermus aquaticus, from which it was originally isolated by chien et al. It consists of a single polypeptide chain with a molecular weight of approximately 95 kda. Protzko and floyd lester erickson department of biological sciences, salisbury university, salisbury, md 21801 abstract.
Pcr protocol for taq dna polymerase with standard taq buffer m0273 protocols. Day 1 start 3ml overnight culture of taq from glycerol stock in lbamp 75ngul day 2 add 1ml of overnight culture to 1l of lbamp. This enzyme has a 5 to 3 dna polymerase exonuclease activity but lacks a 3 to 5 exonuclease activity. Taq is available with different formats to accommodate a variety of pcr applications. Invitrogen accuprime taq dna polymerase, high fidelityformulation with a proofreading enzyme for greater accuracy 9x higher than taq dna polymerase and accuprime accessory proteins for improved speci.
Purification of taq dna polymerase for 1 liter culture modified from the protocol presented in f. Invitrogen taq dna polymerase, recombinant 500 units invitrogen taq dna polymerase, recombinant standard pcr reagents and kits. The higher temperature optimum for the taq polymerase 75c allowed the use of higher temperatures for primer annealing and. Ampliqon taq dna polymerase was compared with a taq dna polymerase from a wellrecognised competitor. Invitrogen taq dna polymerase, recombinant 500 units. Ampliqon taq dna polymerase performed equally well or better in this study. Taq dna polymerase, recombinant protocol 20142taq dna polymerase protocol the example pcr procedure below shows appropriate volumes for a single 50l reaction. Platinum taq dna polymerase is composed of recombinant taq dna polymerase, pyrococcus gb. The optimal amount of taq polymerase to be used per reaction varies with the amplicon length. Gotaq dna polymerase contains gotaq dna polymerase, 5x green gotaq reaction buffer and 5x colorless gotaq reaction buffer.
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