Phoenix Hot Start Taq DNA Polymerase
Frequently Asked Questions
How stable is Phoenix Hot Start Taq when incubated in a PCR reaction mix at room temperature?
Phoenix Hot Start Taq is stable at room temperature for at least 72 hours when incubated in a PCR reaction mix
containing 1X of the supplied, optimized Phoenix Hot Start Taq reaction buffer (standard or GC). Similar testing on a
smaller number of amplicons demonstrated functional stability by endpoint PCR following 20 days room temperature
incubation.
How can PCR cycling conditions be optimized for Phoenix Hot Start Taq?
General Taq protocols for PCR cycling (see recommended cycling conditions from Phoenix Hot Start Taq on Product
Information Sheet) are a good starting point. For determining the optimal temperature for the primer annealing step,
vary the temperature by 2 degree increments starting at about 10 degrees below the Tm of the primer with the lower
Tm up to about 5 degrees above Tm of primer with the higher Tm. Phoenix Hot Start Taq is also compatible with
Touchdown PCR cycling.
Can Phoenix Hot Start Taq utilize cDNA as template for PCR?
Yes.
Is Phoenix Hot Start Taq capable of multiplex PCR?
Yes. Phoenix Hot Start Taq is capable of amplifying multiple amplicons simultaneously in a single 50 µl PCR. Increasing
the amount of polymerase added (up to 2.5 U) may improve results.
How can I optimize Mg2+ conditions for a specific amplicon when using Phoenix Hot Start Taq and the supplied
reaction buffer?
Supplied Phoenix Hot Start Taq buffers contain 2 mM Mg2+ in the final reaction. Therefore, final Mg2+ reaction
concentration may be increased according to user preference with a concentrated solution containing Mg2+. Note
increasing final Mg2+ concentration may lower fidelity of the PCR (1).
When should I use Phoenix Hot Start Taq GC reaction Buffer?
Phoenix Hot Start GC reaction buffer is recommended for use with difficult or GC-rich amplicons (≥ 55%).
100 Cummings Center, Suite 407J, Beverly, MA 01915 • Ph (888) 927-7027 • Fax (978) 867-5724 • www.enzymatics.com
What is the amplification length limit of Phoenix Hot Start Taq?
Phoenix Hot Start Taq has been demonstrated to amplify up to 4 kb human genomic DNA targets, and up to 8 kb lambda
DNA targets.
What is the fidelity/error rate of Phoenix Hot Start Taq?
Phoenix Hot Start Taq has the same fidelity as standard Taq. The fidelity is approximately 2.7 x 10-5 in Phoenix Hot Start
Taq reaction buffer when measured using a LacI-based assay (2).
Why is the Phoenix Hot Start Taq sometimes cloudy upon removing from -20°C storage?
Phoenix Hot Start Taq enzyme solution may appear cloudy immediately upon removing from -20°C degree storage due
the presence of a stabilizer in the storage buffer.
How can yield for long targets be increased when using Phoenix Hot Start Taq?
Use of a PCR enhancer containing Betaine, DTT, BSA and DMSO (3) may be used to improve yield of complex targets ≥ 2
kb.
References:
1. Eckert K. and Kunkel T. A. (1991) Genome Research, 1, 17 - 24.
2. Frey B. and Suppmann B. (1995) Biochemica, 2, 8 - 9.
3. Ralser M., et al. (2006) Biochemical and Biophysical Research Communications, 347, 747 - 751.
Legal Disclaimer:
A portion of this product is licensed from Johnson & Johnson Services, Inc. under U.S. Patent Nos. 5,338,671 and
5,587,287. This product was developed, manufactured, and sold for research use only. Diagnostics rights are available,
please contact Enzymatics for more information.
100 Cummings Center, Suite 407J, Beverly, MA 01915 • Ph (888) 927-7027 • Fax (978) 867-5724 • www.enzymatics.com