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. Innovative qiagen pcr buffer has been developed to save time and effort by reducing the need for pcr optimization. Each promega product is shipped with documentation stating specifications and other technical information. We previously described in a paper published in bios an undergraduate lab activity. T4 dna polymerase certificate of analysis 9pim421 promega. T4 dna polymerase is active in most restriction enzyme buffers and can be used immediately after the restriction enzyme digestion without prior puri. Many of the techniques using polymerases are detailed in promegas protocols and applications guide, third edition, in promega system protocols technical bulletins and technical manuals and in references to these detailed protocols provided within each enzyme description in this guide. Pcr was developed in 19857 and, due to its ability to amplify specific regions of dna. Gotaq dna polymerase contains gotaq dna polymerase, 5x green gotaq reaction buffer and 5x colorless gotaq reaction buffer. Three different dna target varying in length pah 203 bp, eng9 293 bp and eng 5 480 bp were evaluated.
Highperformance taq dna polymerase, nucleotides dntps, buffers and master mixes provide increased reliability and consistency for routine endpoint pcr. Pcr the polymerase chain reaction pcr is a powerful and sensitive technique for dna amplification 1. Pcr products from amplifications with six different taq polymerases af were cloned into pjet1. C 53exonuclease activity no 3 5 exonuclease activity processivity 50 60 nucleotides extension rate 426kb min at 7080. Size units p207b 1,000 p207e 5,000 p407a high conc. Gotaq dna polymerase m300 protocolpdf 207 kb english. Enhanced performance over standard taq dna polymerase. Taq dna polymerase is a thermostable enzyme that synthesizes dna from singlestranded templates in the presence of dntps and a primer. 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. Pcr protocol for taq dna polymerase with standard taq buffer m0273. Polymerase chain reaction pcr background the polymerase chain reaction pcr is the single most commonly used procedure in molecular genetics. Full text full text is available as a scanned copy of the original print version. This enzyme has a 5 3 dna polymerase and a 5 3 exonuclease activity but lacks a. Platinum taq dna polymerase is a recombinant taq polymerase complexed with a proprietary antibody that blocks the polymerase activity at ambient temperatures.
Com number 100 september 2008 7 are removed, and the column flowthroughs are discarded. It is supplied with 10x standard taq reaction buffer, which is detergentfree and designed to be compatible with existing assay systems. Taq dna polymerase from promega and neb were compared to two lots of lucigens econotaq dna polymerase in amplifying the. Sometimes, these matter less, and quick and dirty pcr is all you need. The minicolumns are placed back in the collection tubes, then 200. While most may think standard taq is the backbone of pcr, there are many other dna polymerase options out there. 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. Certificate of analysis gotaq hot start polymerase. Promega notes number 83 2003 21 article subhead abstract gotaq dna polymerase, promegas new formulation of taq dna polymerase, is supplied with two buffersgreen gotaq reaction buffer and colorless gotaq reaction buffer.
Due care should be exercised with all promega products to prevent direct human contact. The evidence of contaminant bacterial dna in several. Ampliqon taq dna polymerase was compared with a taq dna polymerase from a wellrecognised competitor. 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. First isolated from the bacterial thermophile thermus aquaticus, the taq polymerase has become the staple enzyme in molecular biology. Bioneers taq dna polymerase is isolated from recombinant li strain containing the taq dna polymerase gene from thermus aquaticus yt1. Dna polymerase is an enzyme that synthesizes dna molecules from deoxyribonucleotides, the building blocks of dna. Not for use in diagnostic procedures for clinical purposes. Efficiency comparison of seven different taq polymerases. To streamline this process it is helpful to directly load the pcr sample onto an agarose gel without adding gel loading dye or buffer to each sample. Rapid purification of highactivity taq dna polymerase. Thermo scientific taq dna polymerase is a highly thermostable dna polymerase from the thermophilic bacterium thermus aquaticus. Efficiency comparison of seven different taq polymerases used in hemogenetics j. Ampliqon taq dna polymerase performed equally well or better in this study.
Gotaq dna polymerase product information promega corporation. The 5x green gotaq reaction buffer, contains two dyes. The sequence analysis revealed a presence of different bacterial dna table 2 in each taq polymerase. Taq dna polymerase is the industry standard for routine pcr. Recombinant taq dna polymerase purification in todays lab we will purify recombinant taq dna polymerase and assay for its function. If desired, a t4 dna polymerase buffer consisting of 33mm trisacetate ph 7. Commercial producers have sinced cloned and expressed the enzyme in e. Gotaqr flexi dna polymerase product information sheet 9pim829.
Recombinant taq dna polymerase is ideal for standard pcr of amplicons 5 kb or shorter. Our products are backed by a worldwide team of technical support scientists. Gotaq g2 dna polymerase product informationpdf 191 kb english. The 10x buffer supplied with this enzyme has a composition of 250mm trisacetate ph 7. Norgens taq dna polymerase is available with 3 different reaction buffers. Gotaqr qpcr master mix technical manual tm318 promega. Green buffer allows direct gel analysis after amplification.
From each library 20 clones were analyzed and sequenced. Roughly 10 years later, the polymerase chain reaction was developed and shortly thereafter taq became a household word in molecular. Taq dna polymerase thermostable dna polymerase isolated from thermus aquaticus yt1 94 kda protein t 12 40 minutes at 95. Certificate of analysis gotaq flexi dna polymerase.
Onetaq dna polymerase is an optimized blend of taq and deep vent dna polymerases for use with routine and difficult pcr experiments. Links to pubmed are also available for selected references. Dna polymerase econotaq lucigenexcellent results when compared to promega and new england biolabs taq polymerases figure 1. Taq dna polymerase is a highquality recombinant enzyme that is suitable for general and specialized pcr applications see figures tolerance of different primer t m values and specific amplification of long pcr products qiagen pcr buffer. The enzyme catalyzes 53 synthesis of dna, has no detectable 35 exonuclease proofreading activity and possesses low 53 exonuclease activity. The polymerase you use has a significant impact on the efficacy of your pcr, specifically on the product yield, the purity of the product and the faithfulness with which the starting product is transcribed. These enzymes are essential for dna replication and usually work in pairs to create two identical dna strands from a single original dna molecule.
Purification of thermostable dna polymerases gene and. The enzyme is a fulllength form of taq dna polymerase that exhibits 53 exonuclease activity. Taq dna polymerase is a thermostable dna polymerase that catalyzes the polymerization of nucleotides into duplex dna in the 5 3 direction. Activity of t4 dna polymerase in promega s restriction enzyme buffers. Promega products are warranted to meet or exceed the stated specifications. The onetaq reaction buffers and high gc enhancer have been formulated for robust yields with minimal optimization, regardless of a templates gc content onetaq dna polymerase is supplied with two 5x buffers. Currently, str typing is the most efficient and the fastest way for identification of. Efficient cloning and high protein yields the single step krx cells are highly competent,allow bluewhite screening, and have mutations that make them enda, reca,ompt and ompp. All samples were amplified and sscpelectrophoresed in the same experiment. Thermostable polymerases are of course key enablers of molecular biology, due to their usefulness in polymerase chain reaction pcr.
We will discuss who discovered it, the method, and its function. Gotaq dna polymerasea,b is a taq dna polymerase supplied in a proprietary formulation containing 50% glycerol with buffers designed for. It is frequently used in the polymerase chain reaction pcr, a method for greatly amplifying the quantity of short segments of dna. The recombinant protein will be purified from an e.
The enzyme consists of a single polypeptide with a molecular weight of 94 kda. It was found that a fragment of native thermus aquaticus dna polymerase taqwt lacking 288 nterminal amino acids taq. Comparison of the 5 nuclease activities of taq dna. Taq is available with different formats to accommodate a variety of pcr applications. Promega buffer b, promega taq dna polymerase and amplitaq gold buffer c and with amplitaq gold buffer and promega buffer no pcr products obtained. The enzyme is a taq dna polymerase supplied in a proprietary formulation containing 50% glycerol with buffers designed for enhanced amplification.
Get a printable copy pdf file of the complete article 394k, or click on a page image below to browse page by page. Gotaq g2 dna polymerase reliably amplifies a wide range of pcr templates and. Description continued 156t a prepare the gotaq qpcr reaction mix by combining primers, water and gotaq qpcr master mix. Protzko and floyd lester erickson department of biological sciences, salisbury university, salisbury, md 21801 abstract. Janica b a central forensic laboratory of the police, warsaw, poland b department of forensic medicine, medical university of bialystok, poland abstract. During this process, dna polymerase reads the existing dna strands to create two new strands that match the existing ones. The dna polymerase from thermus aquaticus taq polymerase, famous for its use in the polymerase chain reaction, is homologous to escherichia coli dna polymerase i pol i like pol i, taq polymerase has a domain at its amino terminus residues 1290 that has 5 nuclease activity and a domain at its carboxy terminus that catalyses the polymerase reaction. Snp poltaq dna polymerase is a highly selective dna polymerase variant, specially evolved for all assays in which high single nucleotide discrimination is required. Both buffers contain mgcl2 at a concentration of 7. Provided with green and colorless reaction buffers.
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