Bio-Rad iTaq DNA Polymerase User Manual

iTaq™DNA Polymerase

250 U (50 µl) 170-8870 5,000 U (1 ml) 170-8875

For research purposes only Store at -20°C

Storage and Stability

Store the iTaq DNA Polymerase at -20°C in a constant temperature freezer. When stored under these conditions the polymerase is stable for one year after ship date.

Kit Contents

iTaq DNA Polymerase is suitable for many PCR applications. the antibody-mediated hot-start employed by iTaq Polymerase sequesters Taq activity prior to the initial PCR denaturation step. Upon heat activation for three minutes at 95°C, the antibodies denature irreversibly, releasing fully active Taq DNA polymerase. This enables specific and efficient primer extension with the convenience of room temperature reaction assembly.

Reagent Kit Size Volume Description

iTaq DNA polymerase 250 U 50 µl iTaq DNA polymerase, 5 U/µl (Clear Cap) 10X iTaq Buffer 1.25 ml 10X PCR buffer, 200 mM Tris-HCl, (Blue Cap) pH 8.4, 500 mM KCl MgCl

1.25 ml 50 mM MgCl

(Green Cap)

iTaq DNA polymerase 5,000 U 1 ml iTaq DNA polymerase, 5 U/µl (Clear Cap) 10X iTaq Buffer 25 ml 10X PCR buffer, 200 mM Tris-HCl, (Bottle) pH 8.4, 500 mM KCl MgCl

25 ml 50 mM MgCl

(Bottle)

Reaction Set Up

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10X iTaq buffer 5.0 µl 1X 50 mM MgCI

1.5 µl 1.5 mM

10 mM dNTP mix 1.0 µl 200 µM each iTaq DNA Polymerase 0.25 µl 1.25 U Primer 1 x µl 100 nM–500 nM Primer 2 x µl 100 nM–500 nM Sterile water x µl DNA template x µl

Total Volume 50 µl

Typical Thermal Cycling Protocol

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3 min, 95ºC

Polymerase activation

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: 25–40 repeats 30 sec, 95ºC

30 sec, 55ºC 30 sec – X min, 72ºC (depending on length of PCR product)

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Hold at 4ºC

Reagents and Materials Not Supplied

10 mM dNTP mix (Catalog # 170-8874) Microcentrifuge tubes, screw capped Pipette tips, preferably aerosol barrier tips Vortexer Microcentrifuge Tubes for thermal cycler Sterile water Primers iCycler

thermal cycler

Recommendations for Best Results

Due to the sensitivity of PCR, precautions should always be taken to minimize contamination. Steps at all stages of setting up reactions should be designed to prevent reagent contamination with samples, primers, and previous reaction products. This should involve the use of separate areas for reaction set up, template preparation, and reaction product analysis. Also the use of barrier pipet tips and non-flip cap tubes for templates, dilutions, and primers are recommended for reducing contamination.

To learn more about Bio-Rad’s complete solution for amplification, visit our website:

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.

4106202 Rev B

Bio-Rad Laboratories, Inc.

2000 Alfred Nobel Drive, Hercules, CA 94547

510-741-1000