TAQXpedite™ PCR System (FAST End-Point)

Applications

  • Fast PCR with a standard PCR thermocycler.
  • Fast PCR of templates with up to 80% GC content, amplicons longer than 30 kb, and multiplex PCR.

Fast PCR can be achieved using fast PCR instrumentation or by using enzyme/reagent combinations that improve PCR efficiency. With a standard thermocycler, the TAQXpedite™ PCR system achieves fast PCR results with total reaction time as little as 16 minutes for a 500-bp amplicon (Table 1, Fig. 1). The TAQXpedite™ kit can be used with a either a specialized fast PCR thermocycler or a standard thermocycler.

The TAQXpedite PCR System (FAST End-Point) contains a unique blend of thermostable DNA polymerases that can be used for fast PCR, a carefully optimized 2X Universal MasterMix and a Difficult/Long MasterMix with all four dNTPs, and an optimized MgCl2 concentration. The MasterMix also contains Epicentre's patented PCR Enhancer with betaine,* which substantially improves the yield, efficiency, and specificity of amplification of many target sequences, especially those containing a high GC content or secondary structure.1-5 The TAQXpedite PCR System's Universal Master-Mix is designed for routine and/or high-throughput fast PCR applications (Fig. 1), while the Difficult/Long MasterMix is designed for use with troublesome, non-routine, or long applications (Fig. 2). All reaction components are included in the kit except for template and primers.

Benefits

  • Increased sample throughput.
  • High fidelity due to proofreading capabilities.
  • Two convenient MasterMixes for universal and difficult/long amplifications.

Table 1. Total amplification times for TAQXpedite™ and standard PCR protocols.

Amplicon Size TAQXpedite™ Fast Protocol Standard PCR
539 bp 16 min 1 hr 16 min
1 kb 20 min 1 hr 30 min
4 kb 39 min 3 hr 1 min
7 kb 1 hr 39 min 4 hr 31 min
20 kb 2 hr 55 min 11 hr 1 min
30 kb 5 hr 33 min 16 hr 1 min
270 bp high-GC 44 min 1 hr 16 min
Nine-amplicon multiplex 52 min 1 hr 16 min
Figure 1. Fast PCR of a 539-bp amplicon from lambda DNA. Different starting concentrations of template were used in a 16-minute reaction with the TAQXpedite™ kit. Lanes M, 100-bp ladder; lane 1, 100 pg; lane 2, 10 pg; lane 3, 1 pg.
Figure 2. Comparison of the TAQXpedite™ PCR System with competitive kits for amplification of a 4-kb fragment. Gel image depicts results starting with 1 ng lambda DNA as the template for PCR. Lane M, DNA marker; lane 1, TAQXpedite PCR System; lane 2, Competitor Q's Fast Kit; lane 3, Competitor A's Fast PCR MasterMix.

References

  1. Schanke, J.T. and Grunenwald, H.L. (1997) Epicentre Forum 4(1), 2.
  2. Grunenwald, H.L. and Schanke, J.T. (1997) Epicentre Forum 4(1), 4.
  3. Mytelka, D.S. and Chamberlin, M.J. (1996) Nucleic Acids Res. 24, 2774.
  4. Henke, W. et al. (1997) Nucleic Acids Res. 25, 3957.
  5. Weissensteiner, T. and Lanchbury, J.S. (1996) BioTechniques 21, 1102.

TAQXpedite™ PCR System. 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 patent 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.

*Covered by issued and/or pending patents.