Bio-Rad iProof™ High-Fidelity DNA Polymerase User Manual
Iproof, High-fidelity dna polymerase
iProof
™
High-Fidelity DNA Polymerase
2 units/µl, 10 µl
20U
172-5300
2 units/µl, 50 µl
100U
172-5301
2 units/µl, 250 µl
500U
172-5302
For research purposes only
Store at –20°C
iProof is a high-fidelity DNA polymerase that offers extreme performance for all PCR
applications. Incorporating an exciting new and patented technology, iProof DNA
polymerase brings together a novel Pyrococcus-like enzyme with a processivity enhancing
domain. This allows for the generation of long templates with an accuracy and speed
previously unattainable with a single enzyme. The extreme fidelity of iProof makes it a
superior choice for cloning. The error rate of iProof polymerase is determined to be 4.4 x
10
-7
in iProof HF buffer, which is approximately 50-fold lower than that of Thermus
aquaticus, and 6-fold lower than that of Pyrococcus furiosus.
Storage and Stability
Store iProof™ High-Fidelity DNA Polymerase at -20°C in a constant temperature freezer.
When stored under these conditions, the polymerase is stable for one year after the ship
date.
Kit Contents
Reagent
20U
100U
500U
Description
iProof Polymerase
10 µl
50 µl
250 µl
iProof™ High Fidelity DNA
Polymerase, 2 units/µl
iProof HF Buffer
1.5 ml
1.5 ml
4 x 1.5 ml
5X HF Buffer, 7.5 mM MgCl
2
iProof GC Buffer
1.5 ml
1.5 ml
4 x 1.5 ml
5X GC Buffer, 7.5 mM MgCl
2
MgCl
2
1.5 ml
1.5 ml
2 x 1.5 ml
50 mM MgCl
2
solution
DMSO
500 µl
500 µl
500 µl
100% DMSO solution
iProof DNA polymerase is unlike other enzymes. Please read the QuickGuide to
modify your protocol for optimal results.
QuickGuide (See Notes About Cycling Conditions for details)
•
Use 98°C for denaturation.
•
Anneal at T
m
+3°C (>20nt oligo).
•
Use 15–30 sec/kb for extension times. Do not exceed 1 min/kb.
•
Use iProof at 0.5–1.0 U per 50 µl reaction. Do not exceed 2 U/50 µl.
•
Use 200 µM dNTPs. Do not use dUTP.
•
iProof produces blunt end DNA products.
Related Amplification Products From Bio-Rad Laboratories
Reagents for PCR or Real-Time PCR
iProof™ High-Fidelity PCR Kit
172-5331
iProof HF Master Mix
172-5310
iProof GC Master Mix
172-5320
iTaq™ DNA Polymerase
170-8870
iTaq Supermix With ROX
170-8854
iTaq SYBR Green Supermix With ROX
170-8850
iQ™ Supermix
170-8860
iQ SYBR Green Supermix
170-8880
iScript™ cDNA Synthesis Kit
170-8890
iScript Select cDNA Synthesis Kit
170-8896
iScript One-Step RT-PCR Kit with SYBR Green
170-8892
iScript One-Step RT-PCR Kit for Probes
170-8894
For ordering information on larger pack sizes, or to learn more about Bio-Rad amplification
reagents and instruments, visit www.bio-rad.com/amplification/
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 the US Patent 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.
iProof, iTaq, iQ, and iScript are trademarks of Bio-Rad Laboratories..
10002298 Rev B
Bio-Rad Laboratories
2000 Alfred Nobel Drive, Hercules, CA 94547
510-741-1000