Phusion polymerase protocol pdf free

Thermo scientific phusion rtpcr kit product information. In addition, phusion dna polymerases are tolerant of various inhibitors. As a result of this unique functional pairing, phusion dna polymerases are capable of generating pcr products with very high accuracy and speed. All that is required is the addition of template, primers and water. This simplifies the cdna synthesis protocol as it renders a. Place your order before 8pm est for overnight delivery. While straightforward and generally troublefree, there are pitfalls that.

Overlap extension pcr, phusion polymerase or taq polymerase. Phusion highfidelity dna polymerase is comprised of a dnabinding domain fused to a pyrococcuslike proofreading polymerase. Protocol for a routine pcr with phusion highfidelity pcr kit introduction. Due to the high salt concentration in the reaction buffer, phusion dna. Phusion hot start flex dna polymerase is available as standalone enzyme or in a master mix format and enables high specificity amplification of a broad range of templates.

Protocol for phusion highfidelity pcr master mix with gc buffer overview. Manufactured and qualitycontrolled at new england biolabs, thermo scientific phusion highfidelity dna polymerase offers both high fidelity and robust performance. Phusion dna polymerase may be diluted in 1x hf or gc buffer just prior to use in order to reduce pipetting errors. Phusion dna polymerases thermo fisher scientific us. Flow chart of the phusion sitedirected mutagenesis kit protocol. Phusion dna polymerase is an ideal choice for cloning and can be used for long or difficult amplicons. Polymerase, the optimal reaction conditions may differ from pcr protocols for. The following guidelines are provided to ensure successful pcr using phusion dna polymerase. Thermo scientific phusion highfidelity dna polymerase.

Both phusion hf buffer and phusion gc buffer provide 1. Phusion highfidelity dna polymerase is supplied with 5x phusion hf buffer, 5x phusion gc buffer, dmso and 50 mm mgcl2 solution. Phusion the wolfson centre for applied structural biology. Due to the nature of the phusion highfidelity dna polymerase, the optimal reaction conditions may differ from pcr protocols for standard dna polymerases. Download a pdf containing pricing for our full product list. Pcr protocol for phusion highfidelity dna polymerase m0530 protocols. Protocol for a routine pcr with phusion highfidelity pcr. Can i use phusion polymerase in quikchange protocol.

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. As such, conditions recommended below should be used for optimal performance. Phusion highfidelity dna polymerase high performance for. See your thermal cyclers manual to determine the appropriate size tube. Protocol for phusion highfidelity pcr master mix with gc. The following guidelines are provided to ensure successful pcr using phusion master mixes. Phusion highfidelity pcr master mix with hf buffer neb. Im planning to use overlap extension pcr for dna insertion into a vector, as it is explained by briskin and matsmura in. 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. Datasheet for phusion highfidelity dna polymerase neb. Thermo scientific phusion rtpcr kit is a complete kit designed for performing cdna synthesis. Component, 20 l reaction, 50 l reaction, final concentration. In the pcr reaction, phusion dna polymerase extends the primers. Datasheet for phusion hot start flex dna polymerase.

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