Home  Contacts  Order  Catalog  Support  Search  Alphabetical Index  Numerical Index  Restriction Enzymes  Modifying Enzymes  PCR, qPCR, RT-PCR & dNTPs  Molecular Cloning  Nucleic Acid Purification  Molecular Labeling & Detection  In vitro Transcription  Electrophoresis Products  Nucleotides  Transfection Reagents  Reagents C A T A L O G

Long PCR Enzyme Mix*
* Not available in the USA.

#K0181 Supplied with: 10X Long PCR Buffer 10X LOng PCR Buffer with 15 mM MgCl2 MgCl2 Solution DMSO Water, nuclease-free	100 u (5 u/µl)   0.6 ml 0.6 ml 0.6 ml 0.3 ml 1.5 ml
#K0182 Supplied with: 10X Long PCR Buffer 10X LOng PCR Buffer with 15 mM MgCl2 MgCl2 Solution DMSO Water, nuclease-free	500 u (5 u/µl)   2x1.25 ml 2x1.25 ml 2x1.25 ml 1 ml 1.5 ml
Related Documents (in pdf, ~30-60 KB): Certificate of Analysis: #K0181, #K0182 MSDS (English) MSDS (English-USA) MSDS (German) Flyer (448 KB)

 

Features

• Unique blend of two thermostable DNA polymerases.
• Unique long PCR buffer.
• Long PCR products:
  • up to 47 kb with viral DNA as template;
  • up to 21 kb with genomic DNA as template.
• Fidelity is three-times higher than with Taq DNA Polymerase.
• High yield.
• GC-rich templates can be successfully amplified.

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

Description
Long PCR Enzyme Mix is a unique blend of Fermentas Taq DNA Polymerase and a thermostable high fidelity DNA polymerase with proofreading activity. The two enzymes synergistically generate long PCR products with greater yield and fidelity than Taq DNA Polymerase alone. The fidelity of PCR with this enzyme mix is three-times higher than with Taq DNA Polymerase. The ratio of enzymes in the Long PCR Enzyme Mix is optimized for generation of very long amplicons: up to 47 kb with viral DNA and up to 21 kb with genomic DNA templates (see Fig.1). The specially-formulated Long PCR Buffer protects DNA from depurination and nicking during long thermal cycling. The products generated with the Long PCR Enzyme Mix are mostly 3'-dA tailed, and they are suitable for TA cloning.
The Long PCR Enzyme Mix is also used for efficient amplification of GC-rich DNA regions (see Fig.2).

Applications

• Long range PCR.
• PCR from GC-rich or difficult templates.
• Generation of template for TA cloning.
• RT-PCR.

Concentration
5 u/µl

Quality Control
Functionally tested in generation of 47.4 kb amplicon with lambda DNA as template.

Figure 1. Amplification of long DNA fragments.
DNA fragments were amplified from 1.25ng of lambda DNA in 50 µl of reaction mixture containing 2.5 u of Long PCR Enzyme Mix.
M - Lambda Mix Marker
1-5 - PCR products

Figure 2. Successful amplification of GC-rich template with the Long PCR Enzyme Mix.
Human IGFR II gene (85% GC content) was amplified using the Long PCR Enzyme Mix and two other commercial PCR systems for GC-rich templates. PCR mixtures (50 µl) contained 100ng of human genomic DNA, 200 µM each dNTP and 0.5 µM of each primer. Cycling conditions: 94°C, 2 min; 96°C, 10s; 68°C, 1 min 30s; 35 cycles; 72°C, 7 min.
M - GeneRuler™ 100 bp Plus DNA Ladder
1 - Long PCR Enzyme Mix supplemented with 12% DMSO
2-3 - PCR systems for GC-rich templates from other vendors

Related Products

dNTP Mixes dNTP Set InsTAclone™ PCR Cloning Kit 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 Lambda Mix DNA Marker, 19

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 vasario 28, 2008 15:37