High efficient & High fidelity PCR enzyme
KOD FX
(Code.No : KFX-101)
Research Only
TOYOBO CO., LTD. Life Science Department
OSAKA JAPAN
07-05
Code No. KFX-101 200U X 1 Vial
Store at -20℃
High efficient & High fidelity PCR enzyme
KOD FX
KOD FX was developed base on the KOD DNA polymerase from a hyperthermophilic Archaeon Thermococcus kodakaraensis KOD1. KOD FX shows excellent elongation ability, efficiency and reliability in addition to high fidelity. The enzyme solution of KOD FX contains two types of anti-KOD DNA polymerase antibodies inhibit the polymerase and 3’-5’ exonuclease activities, and this enables hot start PCR.
Feature of KOD FX
・High fidelity
The PCR error ratio of KOD FX is about 10 times less than that of rT aq DNA polymerase.
・Excellent elongation ability
KOD FX enables the following amplification (Maximum) : 40kb from lamda phage DNA , 24kb from human genomic DNA, and 13.5kb from cDNA.
・High success rate
This enzyme is effective for the amplification of GC-rich target and crude samples.
１．Contents
reaction)
(200
KOD FX (1.0 U/μl) 200μl × 1 viral
2× PCR Buffer for KOD FX* 1.7ml × 3 vial
2mM dNTPs 1ml × 2 vial
* 2× PCR Buffer for KOD FX is a solution at -20℃.
２．Caution
Research use only.
３．Quality control
The quality check was performed by an amplification of tP A gene (24kb).
４． Protocol
（１） Prepare the reaction mixture as follows;
For 1 reaction
2x PCR buffer for KOD FX 25 μl 1×
明锐pro价格2mM dNTPs
10 μl 0.4mM each 10pmol /μl Primer #1 1.5 μl 0.3μM 10pmol /μl Primer #2
1.5 μl
路肩0.3μM
T emplate DNA
≧ 1 μl
Genomic DNA ～200 ng / 50μl
Plasmid DNA ～50 ng / 50μl
cDNA (from ～200 ng RNA) / 50μl
KOD FX (1.0U/μl) 1 μl 1.0 U / 50μl Autoclaved, distilled water up to 50 μl
（２） Recommended cycle conditions
The following cycling steps are recommended. The step-down cycle condition is effective for the trouble shooting of a smear amplification on a long-distance PCR.f6
2-step cycle
3-step cycle
Predenature : 94℃, 2min. Predenature : 94℃, 2min. Denature : 98℃, 10sec. Denature : 98℃,10sec. Extension : 68℃, 1min. /kb
Annealing : (Tm-5)℃
Extension : 68℃, 1min. /kb
Step-down cycle
Predenature : 94℃，
Extension : 74℃，1min. /kb Denature : 98℃，10sec. Extension : 72℃，1min. /kb Denature : 98℃，10sec. Extension : 70℃，1min. /kb Denature : 98℃，10sec. Extension : 68℃，1min. /kb Extension : 68℃，7min.
・ The appropriate extension time is 1min per 1kb.
25～40
cycles
NOTICE TO PURCHASER: LIMITED LICENSE
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 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 claim (such as the patented 5’ Nuclease Process claims in US Patents Nos. 5,210,015 and 5,487,972), no right to perform any patented method, 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.
The use of betaine in DNA polymerase reactions is covered by intellectual property, including U.S. Patent No.
6,270,962 and related patents issued or pending in other countries ("I.P."), exclusively licensed to EPICENTRE T echnologies Corporation, 726 Post Road, Madison, WI 53713, USA ("EPICENTRE"). The right to use betaine in conjunction with KOD DNA Polymerase in PCR is sublicensed by EPICENTRE to TOYOBO Co., Ltd. solely for research purposes. The purchase of this product conveys to the buyer a limited, non-exclusive, non-transferable right under the I.P. to use the purchased product containing betaine and KOD DNA Polymerase in PCR solely for research purposes. No rights are granted to resell, repackage, or further sublicense. No other license is granted to the buyer, whether expressly, by implication, by estoppel or otherwise.
TOYOBO CO., LTD.
Life Science Department
2-8, Dojima Hama 2-chome, Kita-ku, Osaka 530-8230, JAPAN
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