GE Healthcare
Data file 11-0025-97 AC
Polymerase chain reaction (PCR)
FideliTaq DNA Polymerase
Description
FideliTaq™ DNA Polymerase combines high-quality recombinant
Taq DNA Polymerase with a high-fidelity, proofreading
polymerase. This enzyme blend has the 5’→3’ exonuclease
activity of Taq DNA polymerase as well as the 3’→5’ exonuclease
activity of the proofreading polymerase. This increases
amplification fidelity over Taq DNA Polymerase alone and
allows amplification of longer fragment sizes (1–4). FideliTaq
DNA Polymerase is well suited to PCR-mediated cloning and
long PCR amplification.
M
1
120 kb —
3.0 kb —
2
3
4
M. Marker
1. Numb 455 bp
2. NRAGE 1.55 kb
3. Numb 4.6 kb
4. Lambda 20.7 kb
1.65 kb —
FideliTaq DNA Polymerase offers:
Increased fidelity: For cloning and microarray applications.
500 bp —
Efficiency: Allows amplification of long PCR products.
Compatibility: Generates products whose ends are compatible
with either blunt-end or TA cloning.
Applications
PCR-mediated cloning and long PCR amplification
FideliTaq DNA Polymerase generates PCR products whose ends
are compatible with either blunt-end or TA cloning procedures (5).
Although a range of PCR product sizes can be amplified (Fig 1),
optimization may be necessary for targets longer than 10 kb
from complex templates. For amplification of longer targets
with less optimization and greater yields, use FideliTaq
PCR Master Mix or FideliTaq PCR Master Mix Plus (see
ordering information).
Fig 1. Amplification of diverse targets by FideliTaq DNA Polymerase. Target
(source) Lane 1, Numb 455 bp (10 ng human genomic DNA); lane 2, NRAGE
1.55 kb (10 ng human genomic DNA); lane 3, Numb 4.6 kb (10 ng human
genomic DNA); lane 4 λ target 20.7 kb (10 ng λ DNA). A total of 25 cycles was
used for the λ target and 35 cycles for human genomic DNA targets. A final
concentration of 1.5 mM MgCl2 was used in all reactions.
Components
Enzyme is supplied at a concentration of 5 units/µl in 20 mM
Tris-HCl (pH 8.5), 1 mM DTT, 0.1 mM EDTA, 100 mM KCI, 50%
glycerol, and stabilizers.
10X PCR Buffer: 100 mM Tris-HCl (pH 8.6), 500 mM KCl,
15 mM MgCl2. 25 mM MgCl2 solution.
Note: The 25 mM MgCl2 is provided separately.
Quality control
Functionally tested: FideliTaq DNA Polymerase is tested by
amplifying a 20.7 kb PCR product from λ DNA.
Purity: FideliTaq DNA Polymerase is free of detectable
nonspecific nucleases.
Storage
Store at -20°C
References
RT-PCR
1. B
arnes, W. M. PCR amplification of up to 35-kb DNA with high fidelity and high yield
from lambda bacteriophage templates. Proc. Natl. Acad. Sci. USA 91, 2216–2220 (1994).
2. C
heng, S. et al. Effective amplification of long targets from cloned inserts and human
genomic DNA. Proc. Natl. Acad. Sci. USA 91, 5695–5699 (1994).
3. B
arnes, W. M. The fidelity of Taq polymerase catalyzing PCR is improved by an
N-terminal deletion. Gene 112, 29–35 (1992).
4. C
line, J. et al. PCR fidelity of pfu DNA polymerase and other thermostable DNA
polymerases. Nucleic Acids Res. 24, 3546–3551 (1996).
5. M
agnuson, V. L. et al. Substrate Nucleotide-Determined Non-Templated Addition
of Adenine by Taq DNA Polymerase: Implications for PCR-Based Genotyping and
Cloning. BioTechniques 4, 700–709 (1996).
Ready-To-Go RT-PCR Beads
FideliTaq RT-PCR Master Mix (2X)
RT-PCR Master Mix (2X)
Ordering information
Product
FideliTaq DNA Polymerase
Code number
E71180
Related products
PCR
FideliTaq PCR Master Mix (2X)
E71182
FideliTaq PCR Master Mix Plus
E71183
Taq DNA Polymerase
see catalog for full range
PuReTaq™ Ready-To-Go™ PCR Beads
27-9557-01
(0.2 ml tubes/plate) 96 reactions
PuReTaq Ready-To-Go PCR Beads
27-9557-02
(0.2 ml tubes/plate) 5 x 96 reactions
PuReTaq Ready-To-Go PCR Beads
27-9558-01
(0.5 ml tubes) 100 reactions
PuReTaq Ready-To-Go PCR Beads
27-9559-01
(0.2 ml hinged tube with cap) 96 reactions
illustra™ Hot Start Master Mix
25-1500-01
illustra Hot Start Mix RTG
see catalog for full range
see catalog for full range
E71185
E78370
First-strand synthesis
Product
Code number
First-Strand cDNA Synthesis Kit
Ready-To-Go You-Prime First-Strand Beads
Ready-To-Go T-Primed First-Strand Kit
TimeSaver™ cDNA Synthesis Kit
27-9261-01
27-9264-01
27-9263-01
27-9262-01
Nucleotides
illustra Solution dNTPs
see catalog for full range
mRNA and total RNA Purification
mRNA Purification Kit
mRNA Purification Kit
illustra RNAspin Isolation Kits
QuickPrep™ Micro mRNA Purification Kit
QuickPrep mRNA Purification Kit
27-9258-01
27-9258-02
see catalog for full range
27-9255-01
27-9254-01
DNA purification
ExoSAP-IT™
see catalog for full range
illustra GFX™ PCR DNA and Gel Band Purification Kit
28-9034-70
MicroSpin™ S-400 HR Columns
27-5140-01
100 Base-Pair Ladder
27-4007-01
GE, imagination at work, and GE monogram are trademarks of General Electric Company.
GFX, illustra, MicroSpin, PuReTaq, QuickPrep, Ready-To-Go, and TimeSaver are trademarks
of GE Healthcare companies.
For further information, please visit
www.gelifesciences.com/contact
for your regional representative.
GE Healthcare UK Limited
Amersham Place
Little Chalfont, Buckinghamshire
HP7 9NA, UK
www.gelifesciences.com/illustra
Use of this product is covered by one or more of the following US patents and corresponding
patent claims outside the US: 5,079,352, 5,789,224, 5,618,711, 6,127,155, 5,407,800, 5,322,770,
5,310,652, and claims outside the US corresponding to US Patent No. 4,889,818. The purchase
of this product includes a limited, non-transferable immunity from suit under the foregoing
patent claims for using only this amount of product for the purchaser’s own internal research.
No right under any other patent (such as the patented 5’ Nuclease Process claims in US Patents
Nos. 5,210,015 and 5,487,972) and no right to perform commercial services of any kind,
including without limitation reporting the results of purchaser’s activities for a fee or other
commercial consideration, is conveyed expressly, by implication, or by estoppel. This product
is for research use only. Diagnostic uses under Roche patents require a separate license
from Roche. 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.
All third party trademarks are the property of their respective owners.
© 2006-2008 General Electric Company–All rights reserved. First published June 2006.
All goods and services are sold subject to the terms and conditions of sale of the company
within GE Healthcare which supplies them. A copy of these terms and conditions is available
on request. Contact your local GE Healthcare representative for the most current information.
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GE Healthcare Europe, GmbH, Munzinger Strasse 5, D-79111 Freiburg, Germany.
GE Healthcare Bio-Sciences Corp., 800 Centennial Avenue, P.O. Box 1327, Piscataway, NJ
08855-1327, USA.
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169-0073, Japan.
11-0025-97 AC 07/2008