General description
KOD Hot Start Master Mix is a ready-to-use 2X mixture
optimized for convenient high-fidelity PCR, which utilizes a DNA polymerase
with a low error rate and results in a high degree of accuracy in the
replication of the DNA of interest. The mix contains KOD Hot Start DNA
Polymerase, two monoclonal antibodies, ultra-pure deoxynucleotides, and
reaction buffer with MgSO4. The monoclonal antibodies inhibit the
DNA polymerase and 3′→5′ exonuclease activities at ambient temperatures.
Application
KOD Hot Start Master Mix has been used:
- To
yield PCR product for T7 endonuclease I (T7EI) assay.[1]
- For
PCR amplification of the DNA isolated from Chrysosporium
multifidum, a fungus with antimicrobial activity.[2]
- In
the PCR amplification of cDNA.[3]
- In
two-step PCR for the amplification of the target sequence followed by its
insertion into a vector.[4]
Features and Benefits
- Highest
accuracy, yield, and processivity of commercially available proofreading
DNA polymerases
- Amplifies
genomic DNA templates up to 12 kbp
- Amplifies
plasmid and lambda DNA template up to 21 kbp
- Eliminates
mispriming and primer-dimer formation
- Convenient
ambient-temperature setup compatible with automation
- KOD
HotStart® Buffer ensures optimal PCR performance over a
wide range of targets.
- Offers
high specificity, fidelity and yield during PCR amplification
- Optimized
for ease of use and data reproducibility
- Simplifies
PCR set-up and saves time
- High
consistency with minimal risk of contamination
*Manufactured by Toyobo and distributed by Merck KGaA, Darmstadt, Germany
and/or its affiliates (not in Japan).
Note: 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 patents 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.
Other Notes
- 2 ×
1.25 ml or 10 × 1.25 mL KOD Hot Start Master Mix
References:
Kitabayashi, M., et al. 2002. Biosci. Biotechnol. Biochem.66
(10), 2194. Fujii, S., et al. 1999. J. Mol.
One unit is defined as the amount of enzyme that will
catalyze the incorporation of 10 nmol of dNTP into acid-insoluble form in 30
min at 75°C, in a reaction containing 20 mM Tris-HCl (pH 7.5 at 25°C), 8 mM
MgCl₂, 7.5 mM DTT, 50 µg/ml BSA, 150 µM each of dATP, dCTP, dGTP, dTTP (a mix
of unlabeled and [3H]dTTP), and 150 µg/ml activated calf thymus DNA.
Simply add KOD HotStart® Master Mix to an
equal volume of sample containing DNA template and primers. The final diluted
reaction contains 1U KOD HotStart® DNA Polymerase per 50 μl
reaction. The smaller available size provides sufficient master mix for 100 (50
μl scale), or 250 (20 μl scale) reactions, while the larger size is adequate
for 500 (50 μl scale) or 1250 (20 μl scale) reactions.
Legal Information
* Manufactured by Toyobo and distributed by Merck KGaA,
Darmstadt, Germany and/or its affiliates (not in Japan).Note: 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 patents 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.
HOTSTART is a registered trademark of Molecular BioProducts,
Inc.
NOVAGEN is a registered trademark of Merck KGaA, Darmstadt,
Germany
Disclaimer
Toxicity: Standard Handling (A)
