Toll free in the usa 8003569526 telephone 6082744330 usage information. Promega corporation 2800 woods hollow road madison, wi 537115399 usa telephone 6082744330 toll free 8003569526 fax 6082772516. As a member of the wwpdb, the rcsb pdb curates and annotates pdb data according to agreed upon standards. Taq dna polymerase, native thermo fisher scientific.
Promega m3005 gotaq dna polymerase, 4 x 1ml 5x green gotaq reaction buffer and 4 x 1ml 5x colorless gotaq reaction buffer, mgcl2 at a concentration of 7. Protocol for masteramp taq dna polymerase masteramp taq dna polymerase is from an improved strain of the thermophilic bacterium, thermus aquaticus. It has an extension rate of 35100 ntsec at 75c wittwer 1991 biotechniques 101, 76. Taq polymerase has substantial enzymatic activity at 37c, although its optimal activity is expressed at a much higher temperature approximately 72c. Dyebased and probebased assays for realtime qpcr or rtqpcr. It manufactures reagents including ultrapure nucleotides, dna polymerases and mixes, dna markers, competent cells, products for rna analysis and other general reagents for molecular biology. Dna polymerase duplicates all of its dna strands and the cell passes one copy of each strand to its daughter cell. The recombinant protein will be purified from an e. Nov 25, 1988 full text full text is available as a scanned copy of the original print version. Taq dnapolymerase, from thermus aquaticus recombinant, expressed in escherichia coli catalog number d1806 storage temperature 20 c technical bulletin introduction taq dna polymerase is a thermostable enzyme derived from the thermophilic bacterium thermus aquaticus. Gotaq products offer a choice of taq polymerase formulations for basic. This lowtemperature polymerase activity is the basis for nonspecific amplification associated with mispriming events that occur during the initial phase of the pcr reaction.
The recombinant form of this enzyme is expressed in e. Gotaq green master mix contains two dyes blue and yellow that allow monitoring of progress during electrophoresis. Taq dna polymerase is the most common polymerase used in pcr. The polymerase chain reaction pcr was developed by chemist kary mullis in the 1980s, as a means to make many copies of dna fragments. Vector systems and reagents for easy cloning and expression of pcr products. Native taq dna polymerase with 5x reaction buffers. A simple and efficient method for extraction of taq dna. Toll free in the usa 8003569526 telephone 6082744330 internet. E331, e332, se332, me332, le332 depending on the quantity. Pdf production and evaluation of taq dna polymerase. The enzyme consists of a single polypeptide with a molecular weight of 94 kda. 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. Structure of taq polymerase with dna at the polymerase.
Both buffers contain mgcl2 at a concentration of 7. 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 has greatly facilitated the production and reduced the price of the enzyme. Readytouse master mix format with gotaq dna polymerase. Certificate of analysis gotaq green master mix cat. The 5x green gotaq reaction buffer contains two dyes blue and yellow that separate during electrophoresis to monitor migration progress. Taq polymerase, frequently used for standard pcr applications, exhibits 5. Performance of gotaq g2 hot start polymerase is tested in. Surestart taq dna polymerasegenerally yield the best results. Genscript taq dna polymerases are highly thermostable recombinant dna polymerases and ideally suited for routine pcr reactions. In some cases, such as rapd pcr, adding large volume of general taq dna polymerase 5 u.
The 53 exonuclease activity leaves a 3da overhang on the pcr product, which are convenient for direct ta cloning. Promegas pcr systems, enzymes and reagents are proven in pcr to. Cloning of taq polymeraseamplified pcr products thermo. Mutant taq dna polymerases with improved elongation. Gotaq dna polymerase contains gotaq dna polymerase, 5x green gotaq reaction buffer and 5x colorless gotaq reaction buffer. The extension reaction is typically performed at the optimal temperature for taq dna.
This item has been discontinued by the manufacturer and is no longer available. It gives superior results in a broad range of reaction conditions with most primertemplate pairs but amplification may also tend to an increased unspecifity. Alone, taq dna polymerase is inefficient at amplifying fragments larger than 35kb due to its inability to repair nucleotide. From endpoint pcr to quantitative realtime pcr and rtpcr, we offer a complete portfolio of highquality products to meet your amplification needs. Gotaq dna polymerase is a proprietary formulation of taq polymerase that gives robust amplification equal to and, in some cases, superior to that of standard taq the 5x gotaq green and colorless reaction buffers supplied with gotaq dna polymerase contain mgcl 2 at a concentration of 7.
Allows optimization of mgcl 2 concentration and facilitates directtogel analysis of pcr products. Taq polymerase labeling buffer, taq polymerase jena. Taq dna polymerase with standard taq mgfree buffer neb. Instructions for use of products m8291, m8295, m8296, m8297, m8298.
These molecules are visualized, downloaded, and analyzed by users who range from students to specialized scientists. Taq polymerase, being thermostable, proved ideal for pcr. Highperformance taq dna polymerase, nucleotides dntps, buffers and master. Insertion of the t3 dna polymerase thioredoxin binding. The enzyme is a fulllength form of taq dna polymerase that exhibits 5. The enzyme is a taq dna polymerase supplied in a proprietary formulation containing 50% glycerol with buffers designed for enhanced amplification. Recombinant taq is a 94kda enzyme with an optimal 53 polymerase activity between 7580 c in the presence of a magnesium cofactor. It has a 53 dna polymerase activity and a 53 exonuclease activity see figure. Dna polymerase is used to proofread the original dna strand so that the misplaced base are. It is supplied with 10x standard taq reaction buffer, which is detergentfree and designed to be compatible with existing assay systems. The enzyme is in a recombinant form, expressed in e. Taq polymerase labeling buffer is recommended for dna labeling or mutagenesis. Gotaqr g2 hot start polymerase product information 9pim740. Taq dna polymerase is a thermostable enzyme that synthesizes dna from singlestranded templates in the presence of dntps and a primer.
The master mix is provided as a readytouse, stabilized 2x formulation that includes all components for qpcr except template, primers and probe. Taq dna polymerase from thermus aquaticus sigmaaldrich. Taq dna polymerase is a thermostable dna polymerase that catalyzes the polymerization of nucleotides into duplex dna in the 5 3 direction. Mutants of taq dna polymerase resistant to pcr inhibitors. The enzyme has terminal transferase activity which results in the addition of a single nucleotide adenosine at 3 end of the extension product. A dna polymerase mix was made by combining 5 units of taq dna polymerase and 0. Taq dna polymerase buffered aqueous glycerol solution. Certificate of analysis gotaq flexi dna polymerase. Pdf taq dna polymerase is an enzyme essential in performing polymerase chain reaction pcr which. The swissbased pharmaceutical company hoffmanlaroche is facing a series of legal hearings over the.
Taq dna polymerase is a specialized thermostable enzyme isolated from the thermophilic bacterium thermus aquaticus. Taq dna polymerase storage buffer b article number. Toll free in the usa 8003569526 telephone 6082744330. The buffer system is specially optimized for incorporation of labeled or modified nucleotides into dna. The powerplex16 systemad9,10 is used for human identification. Gotaq dna polymerase, promegas new formulation of taq. Such mixtures contain a mixture dntps required as a substrate for the building of new dna strands, mgcl 2, taq polymerase an enzyme required to building new dna strands, a ph buffer and come mixed in nucleasefree water master mixes for realtime pcr include. M166 application of the substance the mixturelaboratory chemicals details of the supplier of the safety data sheet manufacturersupplier. Gotaqr qpcr master mix technical manual tm318 promega. Taq dna polymerase is a thermostable dna polymerase isolated from an e. Gotaq dna polymerase is a proprietary formulation of taq polymerase that gives robust. The structures to the polymerase domains of taq polymerase and of the klenow fragment kf of pol i are almost identical, whereas the structure of a vestigial editing 35 exonuclease domain of taq polymerase that lies between the other two domains is dramatically altered, resulting in the absence of this activity in the thermostable enzyme. Certificate of analysis gotaq hot start polymerase. The rcsb pdb also provides a variety of tools and resources.
Taq dna polymerase is a thermostable dna polymerase that possesses a 5. It is supplied with 10x standard taq mgfree reaction buffer and mgcl 2. First isolated from the bacterial thermophile thermus aquaticus, the taq polymerase has become the staple enzyme in molecular biology. Users can perform simple and advanced searches based on annotations relating to sequence, structure and function. Recombinant taq dna polymerase purification in todays lab we will purify recombinant taq dna polymerase and assay for its function. A protocol for the use of gotaq dna polymerase with 5x green gotaq reaction buffer and 5x colorless gotaq reaction buffer. A protocol for gotaq probe qpcr master mix, which is optimized for quantitative pcr assays in the hydrolysis probe detection format.
David engelkes lab contains the insert taq polymerase and is published in anal biochem. Set up pcr in less than a minute with convenient master mixes. No ligase, postpcr procedures, or pcr primers containing specific sequences are required. Gotaq dna polymerase is a proprietary formulation of taq dna polymerase that gives robust amplification equal to and, in some cases, superior to that of standard taq dna polymerase for ultimate in convenience, choose gotaq colorless master mix or gotaq green master mix and directly substitute in your current pcr application.
Scientists realized that thermostable heatstable dna polymerases would be needed for pcr to work efficiently. Pyrococcus species gbd polymerase possesses a proofreading ability by virtue of its 3. If you are not completely satisfied with any promega pcr product, we will send a replacement or refund your account. A master mix is a mixture containing precursors and enzymes used as an ingredient in rtpcr techniques in molecular biology. A recent market study published by fmi dna polymerase market. Alleles of str loci are differentiated by the number of copies of the repeat sequence contained within the amplified region and are distinguished from one another using fluorescence detection following electrophoretic separation. Get a printable copy pdf file of the complete article 217k, or click on a page image below to browse page by page. Dna polymerase, is supplied with two buffersgreen gotaq. Includes highperformance gotaq dna polymerase, pcr buffers and master mixes for endpoint pcr, hotstart pcr and longrange pcr. Homemade 2x pcr master mix taq dna polymerase is often purchased as concentrated cocktail mixes that contain the figure 1. Gotaq dna polymerase is a proprietary formulation of taq dna polymerase.
Secondgeneration, highperformance gotaq g2 dna polymerase with mgfree buffers. Bioneers taq dna polymerase is isolated from recombinant li strain containing the dna polymerase gene from thermus aquaticus yt1. Gotaq green master mix is a premixed readytouse solution containing bacterially derived taq dna polymerase, dntps, mgcl 2 and reaction buffers at optimal concentrations for efficient amplification of dna templates by pcr. Promega and roche take up battle over pcr patents nature. Hot start taq dna polymerase is a mixture of taq dna polymerase and an aptamerbased inhibitor. Promega pcr systems, enzymes and reagents are proven in pcr to ensure reliable, highperformance results.
Efficiency comparison of seven different taq polymerases used in. Mytaq dna polymerase is a high performance polymerase that exhibits more robust amplification than other commonly used polymerases, delivering very high yield over a wide range of pcr templates and making it the ideal choice for most pcr assays. Ampliqon taq dna polymerase has a molecular weight of 95 kda and exhibits both a 53 dna polymerase and a 53 exonuclease activity. This 94 kda protein shows no detectable levels of contaminating endonucleases or exonucleases by sdspage. Topo ta cloning provides a highly efficient, 5minute, onestep cloning strategy topo cloning for the direct insertion of taq polymeraseamplified pcr products into a plasmid vector. Pluthero 1993 rapid purification of highactivity taq dna polymerase. The gene that encoded the taq dna polymerase had been cloned and expressed in escherichia coli efficiently. Bioline reagents is a primary manufacturer of specialised molecular biology products for the life science industry and research markets. Taq polymerase for robust pcr with and directtogel convenience. Manual 1 using gotaq dna polymerase with either colorless or green gotaq. This lowtemperature polymerase activity is the basis for nonspecific amplification associated with mispriming events that occur during the initial phase of.
Purification of taq dna polymerase for 1 liter culture modified from the protocol presented in f. Dna polymerase is used to create an extra copy of dna each time the cell divides making less than one mistake in a billion bases. Gotaq dna polymerasea,b is a taq dna polymerase supplied in a proprietary. The sevenyear battle over ownership of rights to the enzyme taq dna polymerase came back to court this. Experience improved pcr performance with gotaq dna polymerase products. Promega corporation 2800 woods hollow road madison, wi 53711 u. Pcr is a most widelyused technique in biology, which utilizes thermostable dna polymerase from thermus aquaticus taq. A protocol for gotaq flexi dna polymerase which provides robust amplification equal to and in some cases superior to conventional taq dna polymerase. Description continued both enzymes are necessary for amplifying long targets.
Latest generation gotaq polymerase highperformance for your everyday pcr needs. It is frequently used in the polymerase chain reaction pcr, a method for greatly amplifying the quantity of short segments of dna. Commercial producers have sinced cloned and expressed the enzyme in e. Platinum taq dna polymerase high fidelity is an enzyme mixture composed of recombinant taq dna polymerase, pyrococcus species gbd polymerase, and platinum taq antibody. M1668taq dna polymerase in storage buffer busen promega. On conducting a thorough research on the historic as well as current growth parameters of dna polymerase market, growth prospects of the market are obtained with maximum. Gotaq dna polymerase product information promega corporation. Promega corporation 2800 woods hollow roadmadison, wi 537115399 u.