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Phusion™ Site-Directed Mutagenesis Kit |
 | | | | Phusion™ Site-Directed Mutagenesis Kit |
| | | Developed & Manufactured By Finnzymes Oy. Distributed by New England Biolabs, Inc. | |
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Prices are in US dollars and valid only for US orders.
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- Licensed for PCR*
- A fast and reliable system for generating mutations
Description:
The Phusion™ Site-Directed Mutagenesis Kit is a versatile and efficient tool for introducing point mutations, insertions or deletions in any type of plasmid DNA. With this kit, the entire plasmid is amplified using phosphorylated primers that introduce the desired changes. The amplified, linear PCR product, containing the desired mutation, is circularized in a 5-minute ligation reaction with Quick T4 DNA Ligase. The resulting plasmid can be then transformed into any competent E. coli cells.
The Phusion Site-Directed Mutagenesis Protocol
Advantages:- Robust and reliable exponential amplification method
- No requirements such as special vectors, restriction sites or methylation status for the target plasmid
- No need to destroy the starting template in a separate step
- Due to the high fidelity of Phusion™ Hot Start DNA Polymerase unwanted secondary mutations are minimized
- Amplification of large plasmids up to 10 kb
- The hot start modification in the polymerase prevents the amplification of non-specific products and unwanted degradation of primers prior to first cycle of PCR
- Quick T4 DNA Ligase included in the kit; no purification steps before or after the ligation
- Compatible with all strains of competent E. coli cells
Kit Components:
Control Plasmid with Control Primer Mix
dNTP Mix
Phusion™ HF Buffer Pack (5X)
Phusion™ Hot Start High-Fidelity DNA Polymerase
Quick Ligation Reaction Buffer (2X)
Quick T4 DNA Ligase
Storage Conditions
Storage Temperature:
-20°C
Stable for at least one year from the packaging date.
FAQs
- How does the Phusion Site-Directed Mutagenesis Kit destroy the wild-type template?
- What plasmid sizes can be amplified using the Phusion Site-Directed Mutagenesis Kit?
- Do I need to purify my plasmid before or after ligation when using the Phusion Site-Directed Mutagenesis Kit?
- What strain should I use for the transformation step when using the Phusion Site-Directed Mutagenesis Kit?
- Are there particular requirements for designing primers to use with the Phusion Site-Directed Mutagenesis Kit?
- What should I use for an annealing temperature with the Phusion Site-Directed Mutagenesis Kit?
- What is the most common reason for incorrect clones when screening mutants?
- Why do I not see any PCR product when using the Phusion Site-Directed Mutagenesis Kit?
- Why is my PCR product missing nucleotides at the ligation site?
- Why is the desired mutation missing from the transformants that I screened?
Reagents Sold Separately
Phusion™ HF Buffer Pack
Phusion™ Hot Start High-Fidelity DNA Polymerase
Companion Products
Phusion™ High-Fidelity DNA Polymerase
Phusion™ Hot Start High-Fidelity DNA Polymerase
Legal
Licenses/Patents/Disclaimers:
* PCR license notice: These products are sold under licensing arrangements of Finnzymes Oy with F. Hoffman-La Roche LTD, Roche Molecular Systems, Inc. and Applied Biosystems. The purchase of these products is accompanied by a limited license to use them in the Polymerase Chain Reaction (PCR) process in conjunction with a thermal cycler whose use in the automated performance of the PCR process is covered by the up-front fee, either by payment to Applied Biosystems or as purchased, i.e. an authorized thermal cycler.
Notice to purchaser: Limited license.
The purchase price of this product includes a limited, non-transferable license under U.S. and foreign patents owned by New England Biolabs, Inc., to use this product. No other license under these patents is conveyed expressly or by implication to the purchaser by the purchase of this product.
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