Thermo scientific taq dna polymerase is a highly thermostable dna polymerase from the thermophilic bacterium thermus aquaticus. In contrast to the normal pcr, only a primer is used, so only linear growth not exponential is observed. Genscript taq dna polymerases are highly thermostable recombinant dna polymerases and ideally suited for routine pcr reactions. Taq dna polymerase is the industry standard for routine pcr. Taq is available with different formats to accommodate a variety of pcr applications. Three different dna target varying in length pah 203 bp, eng9 293 bp and eng 5 480 bp were evaluated. Onetaq dna polymerase versus two leading nonhot start enhanced taq polymerases. Dreamtaq dna polymerase, 5 ul 200 u 500 u 5x500 u 20x500 u 10x dreamtaq bu ffe r 1. Denaturation condition varies depending on the thermal cycler and tubes used for pcr. Optimize routine pcr with this highly thermostable dna polymerase. A 956 bp fragment from human genomic dna was amplified with dreamtaq dna polymerase a and taq dna polymerases from other vendors bf according to manufacturers recommendations using 30 pg, 300 pg, 3 ng and 30 ng of template dna, respectively. Only dreamtaq dna polymerase was able to amplify from all template amounts giving high. Dna fragments of increasing length 160 bp, 345 bp, 727 bp, 1988 bp, 4473 bp, 7500 bp were amplified with dreamtaq dna polymerase a and taq dna polymerases from other vendors bh according to manufacturers recommendations.
The taq dna polymerase is the most successful application of a product derived from an extremophile, with annual sales around half a billion dollars podar and reysenbach, 2006. It is frequently used in the polymerase chain reaction pcr, a method for greatly amplifying the quantity of short segments of dna. Taq functions at higher temperatures than a classic dna polymerase and, in part, even permits better sequencing results, because the gcrich structures can be broken down better. Taq excels at amplifying shorter taq dna polymerase, 1 ul 100 u 500 u 10x taq buffer with kcl 0. Both taq buffers can be used for the same applications. It is supplied with 10x standard taq reaction buffer, which is detergentfree and designed to be compatible with existing assay systems. 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. Ab0192kcl 1 thermo scientific taq dna polymerase with kcl buffer description. Taq dna polymerase is an enzyme essential in performing polymerase chain reaction pcr which has recently become a basic technology in research and diagnostic laboratories. Thermo scientific taq dna polymerase, recombinant taq dna. We will discuss who discovered it, the method, and its function. The enzyme has terminal transferase activity which results in the addition of a single nucleotide adenosine at 3 end of the extension product. Pcr the polymerase chain reaction pcr is a powerful and sensitive technique for dna amplification 1.
Taq dna polymerase, recombinant thermo fisher scientific. The enzyme consists of a single polypeptide with a molecular weight of 94 kda. E331, e332, se332, me332, le332 depending on the quantity. The enzyme is a taq dna polymerase supplied in a proprietary formulation containing 50% glycerol with buffers designed for enhanced amplification. Thermo scientific taq dna polymerase with kcl buffer. Reach the pcr performance of your dreams with thermo scientific dreamtaq reagents that offer. Taq dna polymerases product listing product overview. Dreamtaq dna polymerases thermo fisher scientific us.
Thermo scientific pfu dna polymerase is a highly thermostable dna polymerase from the hyperthermophilic archaeum pyrococcus furiosus. Taq dna polymerase is the original and most commonly used pcr enzyme. The enzyme is supplied with optimized dreamtaq buffer, which includes 20 mm mgcl 2. In conclusion, taq dna polymerase gene has been cloned in our laboratory and can be used for the production of large quantities of this enzyme. Dreamtaq dna polymerases are supplied with specially optimized buffers that enable robust dna amplification with minimal optimization of reaction conditions. The enzyme catalyzes 5 3 synthesis of dna, has no detectable 3 5 exonuclease proofreading activity and possesses low 5 3 exonuclease activity. Amplification of longer amplicons with dreamtaq dna polymerase. This premixed formulation saves time and reduces contamination due to a reduced number of. Taq dna polymerase is a thermostable dna polymerase that possesses a 5. The thermostable enzyme, which was isolated from thermus aquaticus and characterized for the first time, is taq dna polymerase 4. Bioneers taq dna polymerase is isolated from recombinant li strain containing the dna polymerase gene from thermus aquaticus yt1. Only dreamtaq dna polymerase was able to amplify all fragments even up to 7.
The taq dna polymerase from thermus aquaticus is the most commonly used enzyme for in vitro dna amplification using polymerase chain reaction pcr. Dreamtaq dna polymerase uses the same reaction setup and cycling conditions as conventional taq dna polymerase. Dna polymerase facilitate detection of low copy number targets by pcr, j. Pcr master mix is a 2x concentrated solution of taq dna polymerase, dntps, and all of the components required for pcr, except dna template and primers. Taq dna polymerase is a thermostable dna polymerase that catalyzes the polymerization of nucleotides into duplex dna in the 5 3 direction. Features protocol guidelines for preventing contamination. Product information thermo scientific dreamtaq dna polymerase. Purification of thermus aquaticus dna polymerase expressed in escherichia coli.
Certificate of analysis taq hot start dna polymerase. The recombinant protein will be purified from an e. Purification of taq dna polymerase for 1 liter culture modified from the protocol presented in f. No need for a special step to denature the antibody to taq dna polymerase. Pdf the replication fidelities of pfu, taq, vent, deep vent and ultma dna polymerases were compared using a pcrbased forward mutation assay.
Product information thermo scientific dreamtaq dna. Thermo scientific taq dna polymerase, recombinant 5 ul. This enhanced taq dna polymerase is designed to consistently and reliably deliver greater sensitivity, better yields, and longer amplicons. Applications pcr amplification of dna fragments as long as 5 kb 1, see the enclosed protocol. Dna polymerase i from li in the year 1950s, many studies have isolated and characterized a continuum of dna polymerases from both prokaryotic and eukaryotic organisms. Recombinant taq dna polymerase purification in todays lab we will purify recombinant taq dna polymerase and assay for its function. Faststart taq dna polymerase is a versatile enzyme that can be used in a wide variety of applications and on multiple instrument platforms. Themagnesium concentrationdoesnot need to be adjusted. Product information pfu dna polymerase, recombinant. Molecular characterization of thermostable dna polymerase. The enzyme catalyzes the templatedependent polymerization of nucleotides into duplex dna in the 5 3 direction. 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.
Taq polymerase also led to the invention of the pcr machine. Search results for taq dna polymerase at sigmaaldrich. Rapid purification of highactivity taq dna polymerase. Taq polymerase has been employed for some time in sequencing. Neb provides high quality recombinant taq at an exceptional value. Ampliqon taq dna polymerase was compared with a taq dna polymerase from a wellrecognised competitor. The thermo scientific dreamtaq green buffer also supports. 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. Taq dna polymerase is a thermostable enzyme that synthesizes dna from singlestranded templates in the presence of dntps and a primer. Extensive optimization of reaction conditions is not 2required. Pcr protocol for taq dna polymerase with standard taq buffer m0273. Taq dna polymerase buffered aqueous glycerol solution. Amplification and cloning of taq dna polymerase gene from. Thermo scientific dreamtaq dna polymerase is an enhanced taq dna polymerase optimized for all standard pcr applications.
Onetaq dna polymerase is an optimized blend of taq and deep vent r dna polymerases for use with routine and difficult pcr experiments. In this lesson, we will learn about taq polymerase and how it is used to amplify segments of dna in the lab. An ultrapure recombinant thermostable taq dna polymerase obtained by high level expression of the taq dna polymerase gene in e. The presence of the taq dna polymerase gene was confirmed by dna sequencing. Full text full text is available as a scanned copy of the original print version. In order to reduce the cost of research work in thailand, recombinant taq dna polymerase was. Recombinant taq dna polymerase is the ideal tool for standard pcr of templates 5 kb or shorter. Use of this product is covered by one or more of the following us. Cloning, taq dna polymerase, thermus aquaticus introduction taq dna polymerase is an enzyme. It ensures higher sensitivity, longer pcr products and higher yields compared to conventional taq dna polymerase. Ampliqon taq dna polymerase performed equally well or better in this study. The yield and the % purity of the reaction products are shown. Pluthero 1993 rapid purification of highactivity taq dna polymerase. Note recombinant taq dna polymerase is the enzyme of choice for most pcr applications.
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