FideliTaq™ PCR Master Mix (2X)

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 71182  100 reactions (50 µl rctn vol)    $124.00  
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Applications:

  • PCR-mediated cloning
  • Long and accurate PCR amplification
  • High throughput PCR

FideliTaq™ PCR Master Mix (2X) combines Taq DNA Polymerase, a high-fidelity, proofreading polymerase and USB Ultrapure nucleotides in a proprietary buffer formulation. This ready-to-use mix increases amplification fidelity up to 6 times over Taq DNA Polymerase alone and allows for amplification of longer product sizes(1-4). The mix generates products whose ends are compatible with either blunt-end or TA cloning procedures with A-tailed ends favored over blunt ends in an approximately 70:30 ratio(5). FideliTaq PCR Master Mix provides robust and reliable performance for PCR applications in which high-fidelity or longer product sizes are required. Since the mix is pre-formulated, experimental variability is significantly reduced.

Convenient
The pre-mixed formulation saves time and reduces potential contamination errors by eliminating several pipetting steps. For a 50 µl reaction, simply add 25 µl of FideliTaq PCR Master Mix to primers, DNA template and PCR-Qualified H2O. Reactions can be easily performed in 10 µl, 25 µl, 50 µl or 100 µl volumes.

High Fidelity
FideliTaq DNA Polymerase gives up to 6 fold higher fidelity than Taq DNA Polymerase, ideal for cloning and microarray applications.

Increase Product Size and Yield
Amplify short and long PCR products from complex DNA templates with little or no optimization (Fig. 1). For PCR products greater than 2 kb, yields are greatly increased with FideliTaq DNA Polymerase and an enhanced buffer system.

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Fig. 1. Range of targets amplified with FideliTaq™ PCR Master Mix. Single-copy NRAGE, 1.55 kb (Lane 1) and Numb, 7.7 kb (Lane 2) were amplified from 1 ng human genomic DNA. Single-copy β-globin, 23.0 kb (Lane 3) was amplified from 100 ng human genomic DNA. Both the 20.7 kb (Lane 4) and 35.0 kb (Lane 5) lambda targets were amplified from 1 ng lambda DNA. No magnesium optimization was required, as 1.5mM final magnesium concentration was used in all reactions.

Improve Specificity and Sensitivity
Amplify PCR products with low background and from low-copy targets, essential for demanding genomic and cDNA applications with limited sample material (Fig. 2).

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Fig. 2. Sensitivity of FideliTaq™ PCR Master Mix. The single-copy Numb gene was amplified from the indicated amounts of human genomic DNA. Approximately one mammalian cell is represented by 3 pg of genomic DNA.

Stable
The mix withstands repeated freeze-thaw cycles with no observed decrease in performance (Fig. 3).

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Fig. 3. Freeze-thaw stability of FideliTaqTM PCR Master Mix. The master mix was subjected to 15 freeze-thaw cycles alternating between dry ice and room temperature. Following freeze-thaw cycles (F/T), the mix was compared to control mix (C) before treatment. Both short and long targets are shown to demonstrate the robust nature of the mix. Single-copy Numb, 455 bp (Lane 1) was amplified from 30 pg human genomic DNA. β-globin 23 kb target (Lane 2) was amplified from 100 ng human genomic DNA. Lambda 35 kb target (Lane 3) was amplified from 1 ng lambda DNA.

Tested User Friendly™ Functional Test:
Functionally tested in PCR to amplify a 20.7 kb product from lambda DNA.

FideliTaq PCR Master Mix Formulation (2X)
50mM Tris, pH 8.8, 100mM KCl, 3mM MgCl2, 0.4mM dNTPs (dATP, dCTP, dGTP, dTTP), 50 units/ml FideliTaq DNA Polymerase, stabilizers.

Components:
4 x 625 µl FideliTaq PCR Master Mix, 2X
(100 X 50 µl reactions or 200 x 25 µl reactions)
   
Brief Protocol

References:

  1. BARNES, W. M. (1994) Proc. Natl. Acad. Sci. USA 91, pp 2216-2220.
  2. CHENG, S., FOCKLER, C., BARNES, W. M., AND HIGUCHI, R. (1994) Proc. Natl. Acad. Sci. USA 91, pp 5695-5699.
  3. BARNES, W. M. (1992) Gene 112, pp 29-35.
  4. CLINE, J., BRAMAN, J. C., AND HOGREFE, H. H. (1996) Nucleic Acids Res. 24, pp 3546-3551.
  5. MAGNUSON, V. L., ALLY, D. S., NYLUND, S. J., KARANJAWALA, Z. E., RAYMAN, J. B., KNAPP, J. I., LOWE, A. L., GHOSH, S., AND COLLINS, F. S. (1996) BioTechniques 4, pp 700-709.

Storage:
Shipped on dry ice. Store at -20°C. Mix well prior to use.

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