Overview pcr the polymerase chain reaction pcr is a powerful and sensitive technique for dna. Invitrogen taq dna polymerase, rekombinant 3 x 500. One unit of taqdna polymerase is the amount of enzyme required to incorporate 10 nmoles of deoxyribonucleotide into dna. Both will leave a 3 da overhang on 30% of the ends of pcr fragments. Platinum taq dna polymerase2the example pcr procedure below shows appropriate volumes for a single 50l reaction. Taq dna polymerase is the most common polymerase used for. The streamlined, easytofollow protocol provided with the kit simplifies pcr setup. One example of this is platinum taq polymerase from invitrogen life. 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 polymerase has been employed for some time in sequencing. Takara taq dna polymerase is a recombinantversion taq polymerase derived from the thermus aquaticus yt1 strain, and is suitable for routine pcr applications. Pcr protocol for taq dna polymerase with standard taq buffer m0273 overview. The polymerase chain reaction pcr is a powerful and sensitive technique for dna amplification.
Taq dna polymerase is a thermostable dna polymerase isolated from an e. Pcr protocol for taq dna polymerase with standard taq buffer m0273 protocols. 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. A simple and efficient method for extraction of taq dna. Protocol the procedure on the following page is suggested as a guideline and starting point when using platinum. Taq dna polymerase catalyzes the incorporation of dntps into dna. Taq dna polymerase is a highly thermostable dna polymerase of the thermophilic bacterium thermus aquaticus. The enzyme is a taq dna polymerase supplied in a proprietary formulation containing 50% glycerol with buffers designed for enhanced amplification. Invitrogen taq dna polymerase, recombinant nondistribution product as customer accommodation. Pcr protocol for taq dna polymerase with standard taq. Invitrogen taq dna polymerase, rekombinant 3 x 500 einheiten invitrogen taq dna polymerase, rekombinant standardpcrreagenzien. Taq dna polymerase is supplied with the unique qiagen pcr buffer that. The streamlined, easytofollow protocol provided with the kit. Hot start taq polymerase invitrogen taq polymerase.
Neb provides high quality recombinant taq at an exceptional value. 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. Platinum taq dna polymerase thermo fisher scientific. Taq dna polymerase is an enzyme widely used in pcr 2.
Taq dna polymerase pcr buffer thermo fisher scientific. We offer a wide variety of pcr enzymes, master mixes, and pcr protocols to meet your experimental needs for routine pcr, qpcr, or rtpcr. Taq dna polymerase with standard taq buffer new england. Ensure successful pcr using nebs pcr protocol for taq dna polymerase with standard taq buffer m0273. Efficiency comparison of seven different taq polymerases. Add all components as in the basic pcr protocol, except for the taq dna polymerase. It is supplied with 10x standard taq reaction buffer. The invitrogen platinum pfx dna polymerase is a proprietary enzyme preparation containing recombinant dna polymerase from thermococcus species kod.
Available on gsava contract for federal government. Optimal reaction conditions incubation times and temperatures, concentration of platinum. Source recombinant enzyme is purified from the cloned thermus aquaticus dna polymerase gene. Dna polymerase is supplied with a streamlined, optimized protocol for fast and easy. Hot start taq dna polymerase for highly specific and sensitive amplification using pcr. Hotstartaq dna polymerase, a modified form of taq dna polymerase. The 53 exonuclease activity leaves a 3da overhang on. Pfx50 dna polymerase offers 50 times better fidelity than taq dna polymerase, coupled with high specificity and an extremely fast elongation rate as fast as 15 seconds per kb. Purification of taq dna polymerase for 1 liter culture modified from the protocol presented in f. Optimal reaction conditions incubation times and temperatures, concentration of taq dna polymerase, primers, mgcl 2, and template dna. The recombinant protein will be purified from an e. In contrast to the normal pcr, only a primer is used, so only linear growth not exponential is observed. Pcr the polymerase chain reaction pcr is a powerful and sensitive technique for dna amplification 1.
Superscript iii firststrand synthesis system for rtpcr. Switch to thermo scientific dreamtaq dna polymerases for better results and added convenience. Taq dna polymerase, recombinant protocol 20142taq dna polymerase protocol the example pcr procedure below shows appropriate volumes for a single 50l reaction. Pluthero 1993 rapid purification of highactivity taq dna polymerase. The trademarks mentioned herein are the property of life technologies corporation andor its affiliates or their respective owners. The following guidelines are provided to ensure successful pcr using nebs taq dna polymerase. Pfuturbo dna polymerase can be used to amplify complex genomic dna targets up to 19 kb and vector targets up to 15 kb in length. Taq is available with different formats to accommodate a variety of pcr applications.
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 dna polymerase is a highly thermostable recombinant dna polymerase of the thermophilic bacterium thermus aquaticus. Taq with standard taq buffer is available in economical extralarge pack sizes. Invitrogen sells the native form of the taq dna polymerase and a cloned version that is expressed in e. The polymerase chain reaction pcr is a powerful and sensitive technique for dna amplification 1. Platinum pfx dna polymerase product information sheet. Invitrogen taq dna polymerase, recombinant 500 units. Taq dna polymerase ensures highly specific pcr for a range of applications with minimal optimization of pcr parameters. Taq dna polymerase, recombinant thermo fisher scientific. Taq dna polymerase taq hot start dna polymerase onetaq dna polymerase is supplied with two 5x buffers standard and gc, as well as a high gc enhancer solution. Prevent the extension of nonspecifically bound primers using this hot start enzyme. The activity of the purified taq polymerase was determined by titration against commercial preparations in a dna. The enzyme is a taq dna polymerase with buffers designed for.
In general pfuturbo dna polymerase amplifies complex targets in higher yield than taq dna polymerase or other commercially available proofreading pcr enzymes. Recombinant taq dna polymerase purification in todays lab we will purify recombinant taq dna polymerase and assay for its function. Taq polymerase can be chemically inactivated at low temperatures, and this modification can be reversed at a high temperature. Taq dna polymerase is the industry standard for routine pcr. A protocol for the use of gotaq dna polymerase with 5x green gotaq reaction buffer and 5x colorless gotaq reaction buffer. Rapid purification of highactivity taq dna polymerase. For multiple reactions, prepare a master mix of components common to all reactions to minimize pipetting error, then dispense appropriate volumes into each 0.
The recombinant taq polymerase, named amplitaq dna polymerase, was commonly used. Accuprime taq dna polymerase, high fidelity, product. Pcr is a most widelyused technique in biology, which utilizes thermostable dna polymerase from thermus aquaticus taq. For multiple reactions, prepare a master mix of components common to all reactions to minimize pipetting error, and then dispense appropriate volumes into each 0. One unit of taqdna polymerase is the amount of enzyme required to incorporate 10 nmoles of deoxyribonucleotide into dna in 30 minutes at 74c. Taq dna polymerase exhibits a very useful habit of adding an extra nucleotide, most often. The invitrogen accuprime taq dna polymerase, high fidelity, provides qualified reagents for the highfidelity pcr amplification of nucleic acid templates.
For most routine andor at rich amplicons or complex invitrogen platinum taq 302, 94c ab read content thermus aquaticus wikipedia. Best profiles from samples were given by taq dna invitrogen, amplitaq gold, jumpstart taq and gel form polymerases. The enzyme is a taq dna polymerase with buffers designed for enhanced amplification. Pcr optimization the choice of the pcr enzyme in combination with an appropriate buffer can. Source recombinant enzyme is purified from the cloned thermus aquaticus dna polymerase gene expressed in e. Taq dna polymerase, primers, mgcl 2, and template dna. Reid phd, in molecular protocols in transfusion medicine, 2000.