Phusion highfidelity dna polymerase, new england biolabs. Choose phusion polymerase from thermo for highly accurate, fast and robust pcr. Elmer 480 thermal cycler manual protocol and numerous book collections from fictions to scientific research in any way. Manufactured and qualitycontrolled at new england biolabs, thermo scientific phusion highfidelity dna polymerase offers both high fidelity and robust performance. Thermo scientific phusion dnapolymerase met hoge getrouwheid 500u. Other phusion hot start flex dna polymerase product introduction for high speed and high performance pcr. Phusion highfidelity pcr master mix with hf buffer is a 2x master mix consisting of phusion dna polymerase, deoxynucleotides and reaction buffer that has been optimized and includes mgcl 2. Thermo scientific phusion green hot start ii highfidelity. This is the pcr protocol for phusion highfidelity dna polymerase m0530. Phusion highfidelity dna polymerase is comprised of a dnabinding domain fused to a pyrococcuslike proofreading polymerase.
Thermo scientific phusion hot start ii high fidelity dna polymerase is the most accurate hot start dna polymerase on the market. Both the dna polymerase and the proofreading activities of phusion hot start ii dna polymerase are inactivated at room temperature by a reversibly binding affibody ligand. Any remaining phusion hot start ii dna polymerase will degrade the a overhangs, creating blunt ends again. Phusion was developed by finnzymes oy, now a part of thermo fisher scientific. Affibody inhibits the dna polymerase activity at room temperature and thus prevents the amplification of nonspecific products. Phusion dna polymerases thermo fisher scientific fr.
Overlap extension pcr, phusion polymerase or taq polymerase. New england biolabs is pleased to announce that it will continue to provide its customers with phusion dna polymerase and related products, which have been distributed for the last six years on behalf of finnzymes oy. Due to the unique nature of phusion dna polymerases, please pay special attention to the guidelines listed below. Product information pfu dna polymerase, recombinant. Phusion hot start ii highfidelity dna polymerase is capable of amplifying long amplicons such as the 7. Phusion highfidelity dna polymerase high performance for. Dna polymerase 1, phusion is one of the most accurate. First isolated from the bacterial thermophile thermus aquaticus, the taq polymerase has become the staple enzyme in molecular biology. Both new england biolabs and thermo fisher scientific offer phusionbased sitedirected mutagenesis kits, which use nonoverlapping primers for pcr, and the pcr products are ligated by dna ligase before transformation.
Product information thermo scientific phusion site. New insights into the quikchangetm process guide the use of. Thermo scientific phusion sitedirected mutagenesis kit is a versatile and efficient tool for introducing point mutations, insertions, or deletions in any type of plasmid dna. Pcr product with thermo scientific dna polymerase, for taq example. In the first step, genomic dna is amplified directly from whole blood using thermo scientific phusion blood direct pcr kit. Thermo scientific phusion highfidelity dna polymerase 2 u. Phusion highfidelity dna polymerase thermo fisher scientific. Jan 30, 2015 phusion offers several advantages over pfuturbo, being faster, more robust and with higher fidelity. Thermo scientific f530s phusion highfidelity dna polymerase is supplied with 5x phusion hf buffer, 5x phusion gc buffer, dmso and 50 mm mgcl2 solution. A separate tube of dmso is provided with the product. All that is required is the addition of template, primers and water. Dna used in thermo fisher scientific quality control assays. Product information thermo scientific phusion sitedirected. Phusion hot start ii dna polymerase possesses the the optimal annealing temperature for phusion hot start ii dna polymerase may differ significantly from that of taqbased access doc.
Thermo scientific phusion highfidelity dna polymerase offers extreme performance for all major pcr applications. This product is manufactured by new england biolabs, inc. Phusion dna polymerases thermo fisher scientific us. With this kit, the entire plasmid is amplified using phosphorylated primers that introduce the desired changes. It combines the phusion dna polymerase and a reversibly bound, specific affibody ligand. Note thermo scientific phusion blood direct pcr kit. Phusion hot start ii dna polymerase combines the dna polymerase and a reversibly bound, specific affibody protein. Recombinant taq is a 94kda enzyme with an optimal 53 polymerase activity between 7580 c in the presence of a magnesium cofactor. Incorporating an exciting technology, phusion dna polymerase brings together a novel pyrococcuslike enzyme with a processivityenhancing domain.
Shop online for a wide selection of thermo scientific phusion highfidelity dna polymerase promotion get superior performance for cloning and other applications requiring. Thermo scientific phusion green highfidelity dna polymerase. Thermo scientific f530l phusion highfidelity dna polymerase is supplied with 5x phusion hf buffer, 5x phusion gc buffer, dmso and 50 mm mgcl2 solution. Phusion dna polymerase is an ideal choice for cloning and can be used for long or difficult amplicons. Pcr amplifies specific dna sequences exponentially by using multiple cycles of a threestep process. Thermo scientific phusion highfidelity pcr kit fisher. The doublestrand dna binding domain blue is fused together with a novel pyrococcuslike enzyme green forming a unique high performance polymerase phusion dna polymerase. The phusion dna polymerase generates long templates. For hot start pcr, thermo scientific tm phusion hot start ii highfidelity dna polymerase is an ideal choice allowing extreme specificity and improved robustness.
Its unique structure, a novel pyrococcuslike enzyme fused with a processivityenhancing domain, increases fidelity and speed. Dna polymerases with high fidelity are important for applications in which the dna sequence needs to be correct after amplification. Both phusion hf buffer and phusion gc buffer provide 1. Manufactured and qualitycontrolled at new england biolabs, thermo scientific phusion highfidelity. The enzyme catalyzes the templatedependent polymerization of nucleotides into duplex dna in the 5 3 direction. Phusion dna polymerase was developed by finnzymes oy, now a part of thermo fisher scientific. You can contact me personally for an improved protocol if you want. The following guidelines are provided to ensure successful pcr using phusion dna polymerase. This kit uses the highly processive thermo scientific phusion hot start ii highfidelity dna polymerase for exponential pcr amplification of dsdna plasmid to be mutated.
Thermo scientific phusion hot start ii highfidelity pcr. Includes phusion dna polymerase 2 ul 5x phusion hf buffer 5x phusion gc buffer dmso 50 mm mgcl 2 solution both phusion hf buffer and phusion gc buffer provide 1. Phusion dna polymerase may be diluted in 1x hf or gc buffer just. The thermo scientific phusion highfidelity dna polymerase offers high performance for all major pcr applications. Thermo scientific phusion highfidelity pcr kit 125 x 20ul rxns get superior performance for cloning and other applications requiring high fidelity with one of the most accurate thermostable dna polymerases available. The polymerase chain reaction pcr is a powerful and sensitive technique for dna amplification. Manufactured and qualitycontrolled at new england biolabs, thermo scientific phusion highfidelity dna polymerase offers both high fidelity and robust performance, and thus can be used for all pcr applications.
Phusion highfidelity dna polymerase for high speed and high performance pcr. Any remaining phusion dna polymerase will degrade the a overhangs, creating blunt ends again. Pcr protocol for phusion highfidelity dna polymerase m0530 protocols. New england biolabs is pleased to announce that it will continue to provide its customers with phusion dna polymerase and related products, which have been distributed for. Phusion dna polymerases offer robust performance with short protocol times. Product information thermo scientific phusion highfidelity. Product information pfu dna polymerase, recombinant pfu. Phusion dna polymerases from thermo fisher scientific description one of the highest referenced polymerase lines in thousands of publications, the thermo scientific phusion highfidelity polymerases are highly accurate and robust products that provide high. Thermo scientific phusion sitedirected mutagenesis kit, point mutations, insertions and deletions can be introduced in any type of plasmid dna. Thermo scientific phusion hot start ii dna polymerase 2 u.
Thermo scientific phusion highfidelity dna polymerases set a gold. The structure of phusion highfidelity dna polymerase. Product information thermo scientific phusion high. New insights into the quikchangetm process guide the use. Protocol for a routine pcr with phusion highfidelity pcr. Phusion highfidelity pcr master mix is convenient 2x mix containing phusion dna polymerase, nucleotides and optimized reaction buffer including mgcl 2. Datasheet for phusion highfidelity dna polymerase neb. Phusion is a registered trademark and property of thermo fisher scientific. Phusion hot start ii dna polymerase does not require a separate activation step in the pcr protocol as it is immediately reactivated at high temperatures. Thermo scientific phusion highfidelity dna polymerase promotion. Phusion highfidelity dna polymerase for high speed and high fidelity pcr, 52x greater fidelity than taq. Thermo scientific phusion highfidelity dna polymerases offer extreme. Commercial producers have sinced cloned and expressed the enzyme in e.
Phusion highfidelity dna polymerase new england biolabs. Taq polymerase from thermo fisher scientific biocompare. Phusion offers several advantages over pfuturbo, being faster, more robust and with higher fidelity. The phusion highfidelity dna polymerase brings together a pyrococcuslike enzyme with a processivityenhancing domain. Phusion dna polymerases offer robust performance with short protocol times, even in the presence of pcr inhibitors, and generate higher yields with lower enzyme amounts than other dna polymerase.
The enzyme, originally developed by finnzymes, is a fusion of a proofreading polymerase and a processivityenhancing domain, creating one of the most reliable and robust polymerases available. Phusion dna polymerases from thermo fisher scientific description one of the highest referenced polymerase lines in thousands of publications, the thermo scientific phusion highfidelity polymerases are highly accurate and robust products that provide high yields with short reaction times and enhanced specificity. England biolabs, thermo scientific phusion highfidelity dna polymerase offers both high fidelity. Product information thermo scientific phusion hot start ii. I have tried 20 different annealing temperatures from 50 to 70 deg c, i have even tried a 2 step pcr skipped the annealing step as the tm was 72 deg c with phusion polymerase and 5x hf buffer. Both new england biolabs and thermo fisher scientific offer phusion based sitedirected mutagenesis kits, which use nonoverlapping primers for pcr, and the pcr products are ligated by dna ligase before transformation. For the 2step protocol, annealing and extansion are performed at the same temperature. Thermo scientific phusion hot start ii dna polymerase 2 ul. However, before adding the overhangs it is very important to remove all phusion hot start ii dna polymerase by purifying the pcr product carefully. Thermo scientific pfu dna polymerase is a highly thermostable dna polymerase from the hyperthermophilic archaeum pyrococcus furiosus. Other phusion highfidelity dna polymerase product introduction for high speed and high performance pcr.
Phusion green hot start ii highfidelity dna polymerase. Since their introduction, thermo scientific phusion highfidelity dna polymerases have been referenced in thousands of publications for highperformance pcr and have become the choice for a multitude of demanding applications ranging from reconstruction, design and massivelyparallel, highthroughput sequencing of whole genomes. Any any remaining phusion highfidelity dna polymerase will degrade the a overhangs, creating blunt ends again. Phusion highfidelity dna polymerase 2 ul thermo fisher. Phusion dna polymerases offer robust performance with short protocol times, even in the presence of pcr inhibitors, and generate higher yields with lower.
Phusion dna polymerases from thermo fisher scientific. I tried to amplify my pcr product which has a size of 1800 bases by using phusion dna polymerase enz. This application protocol describes how pcrrflp restriction fragment length polymorphism assays can be performed from blood samples without dna purification. Thermo scientific phusion highfidelity dna polymerase. Phusion highfidelity dna polymerase produce blunt end dna products. Phusion blood direct pcr kit from thermo fisher scientific. Phusion dna polymerases are tolerant of various inhibitors allowing for robust amplification of pcr products with minimal optimization. Book perkin elmer 480 thermal cycler manual protocol. The unique phusion highfidelity dna polymerase offers robust performance and can be used for all pcr applications. Use the neb tm calculator to estimate an appropriate annealing temperature when using neb pcr products. Thermo scientific phusion highfidelity dna polymerases set a gold standard. New phusion green hot start ii highfidelity dna polymerase is a combination of phusion hot start ii dna polymerase and 5x phusion green buffers. Phusion highfidelity dna polymerase high performance for all.
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