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High Fidelity PCR Enzyme Mix*
* Not available in the USA.

#K0191 Supplied with: 10X High Fidelity PCR Buffer 10X High Fidelity PCR Buffer with 15 mM MgCl2 MgCl2 Solution Water, nuclease-free	100 u (5 u/µl)   0.6 ml 0.6 ml 0.6 ml 1.5 ml
#K0192 Supplied with: 10X High Fidelity PCR Buffer 10X High Fidelity PCR Buffer with 15 mM MgCl2 MgCl2 Solution Water, nuclease-free	500 u (5 u/µl) 2x1.25 ml 2x1.25 ml 2x1.25 ml 1.5 ml
Related Documents (in pdf, ~30-60 KB): Certificate of Analysis: #K0191, #K0192 MSDS (English) MSDS (English-USA) MSDS (German) Flyer (448 KB)

 

Features

• Unique blend of two thermostable DNA polymerases.
• Four-times greater accuracy than with Taq DNA Polymerase.
• Long amplification products:
  • up to 20 kb with viral DNA as template;
  • up to 10 kb with genomic DNA as template.
• High yields.
• Increased efficiency.

Enzyme Mixes for Long and Accurate PCR. Pfu DNA Polymerase, flyer in pdf, 448 KB

Description
The High Fidelity PCR Enzyme Mix is a unique blend of Fermentas Taq DNA Polymerase and a thermostable DNA polymerase with proofreading activity. The two enzymes synergistically generate longer PCR products with greater yield and fidelity than Taq DNA Polymerase alone. Thus, fidelity of the High Fidelity PCR Enzyme Mix exceeds more than four-times the fidelity of Taq DNA Polymerase. In addition, the enzyme mix can generate long amplicons, which are up to 20 kb with viral DNA templates and up to 10 kb with genomic DNA templates. Such long amplicons are often difficult to generate with a single DNA polymerase.

Applications

• Generation of PCR products with high fidelity for purposes of cloning, expression, site-directed mutagenesis.
• Long range PCR.
• RT-PCR.
• Production of higher yields of PCR products.

Concentration
5 u/µl

Quality Control
Generation of 20 kb and 9 kb amplicons with lambda DNA and human DNA as templates, respectively.

Note  

• The High Fidelity PCR Enzyme Mix generates both blunt-ended and 3'-dA tailed PCR products.
• PCR product blunting prior cloning is highly recommended.
• For efficient PCR product cloning use CloneJET™ PCR Cloning Kit.

	M - GeneRuler™ DNA Ladder Mix 1 - 0.3 u of enzyme or enzyme mix 2 - 0.6 u of enzyme or enzyme mix 3 - 1.25 u of enzyme or enzyme mix 4 - 2.5 u of enzyme or enzyme mix 5 - 5 u of enzyme or enzyme mix
Amplification of single copy gene from human genomic DNA using Fermentas Taq DNA Polymerase (recombinant), Pfu DNA Polymerase (recombinant),  Long PCR Enzyme Mix and High Fidelity PCR Enzyme Mix. DNA fragment was amplified from 0.2 µg of human genomic DNA in 100 µl of reaction mixture.
Accuracy of thermostable DNA polymerases and DNA polymerase mixes. Accuracy - an average number of nucleotides the polymerase incorporates before making an error. Polymerase accuracy was measured by amplification of a 323 bp lacZ a gene and analysis of the resulting cloned PCR products using the modified method (1).

Related Products

dNTP Mixes dNTP Set T4 DNA Polymerase FastRuler™ DNA Ladders, ready-to-use CloneJET™ PCR Cloning Kit Genomic DNA Purification Kit DNA Gel Extraction Kit GeneJET™ Plasmid Miniprep Kit First Strand cDNA Synthesis Kit RevertAid™ First Strand cDNA Synthesis Kits Agarase Water, nuclease-free

Reference

1. Lundberg, K.S., et al., High-fidelity amplification using a thermostable DNA polymerase isolated from Pyrococcus furiosus, Gene, 108, 1-6, 1991.

Purchase of this product includes an immunity from suit under patents specified in the product insert to use only the amount purchased for the purchaser’s own internal research. No other patent rights  (such as 5' Nuclease Process patent rights) are conveyed expressly, by implication, or by estoppel.  Further information on purchasing licenses may be obtained by contacting the Director of Licensing, Applied Biosystems, 850 Lincoln Centre Drive, Foster City, California 94404, USA. For complete license disclaimer, see.

Updated balandžio 24, 2008 19:11