Product Datasheet
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| Product Description |
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| Product: |
PyFi High Fidelity DNA Polymerase |
| Catalog Number: |
PYFI |
| Product Page: |
https://www.mol-innov.com/products/ |
| Description: |
PyFi High Fidelity DNA Polymerase is a genetically modified recombinant thermostable enzyme cloned from Pyrococcus furiosus with increased processivity and doubled accuracy compared to standard Pfu polymerase. PyFi contains an integral 3'-5' proofreading capability that increases the fidelity to 50X that of Taq DNA polymerase. PyFi is more sensitive than some leading competitors and creates products with blunt ends. Suitable for Standard Endpoint PCR, Blunt End Cloning, and Mutagenesis of DNA templates up to 8.5kb. PyFi undergoes extensive quality control and is free of contaminating endonucleases. Includes 10X reaction buffer.
Reaction setup: The following setup is recommended for a 50µl reaction but can vary depending on the template and primers being used.
| Component | Volume | Final Concentration |
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| 10X Reaction Buffer | 5µl | 1X | | dNTP, 10mM | 2µl | 400µM | | 5' Primer, 10µM | 1µl | 0.2µM | | 3' Primer, 10µM | 1µl | 0.2µM | | DNA Template | 0.1-5µl | 1-100ng | | PyFi* | 0.5µl | 1.25U | | PCR Grade Water | QS to 50µl | *Add enzyme as last component.
Thermal cycling conditions: The following general cycling conditions are recommended but can vary depending on the template and primers being used.
Three-step protocol | Cycling Step | Temperature | Holding Time | Cycles |
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| Initial Denaturation | 95°C | 1-2min | 1 | | Denaturation | 95°C | 15sec | 25-30 | | Annealing* | 50-68°C | 20-30sec | | Extension | 68-72°C | 1min per kb | | Final Extension | 68°C | 1min per kb | 1 | *Annealing temperature will depend on primer length and composition. Generally, begin 5°C below primer Tm.
Two-step protocol for amplification of >3kb fragments | Cycling Step | Temperature | Holding Time | Cycles |
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| Initial Denaturation | 95°C | 2min | 1 | | Denaturation | 95°C | 20sec | 25-30 | Annealing /
Extension | 68°C | 30sec per kb | | Final Extension | 68°C | 30sec per kb | 1 | |
| Recommended Storage: |
SAMPLE |
| Minimum Shelf Life: |
180.50 |
| Source: |
2.5 U/ul |
| Molecular Weight: |
Proteins |
| Antigen: |
-20 C |
| Target: |
Liquid |
| Target Molecular Weight: |
MOLECULAR BIOLOGY REAGENTS |
| Host: |
351.5 |
| Clonality: |
5 |
| Isotype: |
E. coli |
| Applications: |
Dry ice |
| Concentration: |
190 |
| Volume: |
370 |
| Titer: |
0.02M Tris-HCl; 0.1mM EDTA; 1mM DTT; Tween-20; Nonidet P-40; 50% Glycerol; pH 8.0 |
| Buffer: |
January, 2016 |
| Form: |
200 units |
| Preservative: |
0.08 ml |
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| Optimal experimental conditions and working dilutions must be determined by the end user. Sample concentration and volume are for example purposes only and do not necessarily represent the product that will be received. Please contact us for a current lot-specific product datasheet. |
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| For in vitro laboratory use only |
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Molecular Innovations, Inc. - 46430 Peary Court, Novi, MI 48377, USA
Tel: (888) 557-5055 - Fax: (248) 896-0148 - info@mol-innov.com |