FideliTaq™ DNA Polymerase

 TUF
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 71180 50 units    $39.00  
  250 units    $166.00  
  1000 units    $599.00  
  5 x 250 units    $729.00  
  5000 units    $2,573.00  
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Source: Recombinant proteins expressed in E. coli.

Applications:

  • PCR-mediated cloning
  • Long and accurate PCR amplification

Description:
FideliTaq™ DNA Polymerase combines high quality USB recombinant Taq DNA Polymerase with a high-fidelity, proofreading polymerase. This enzyme blend has the 5’3’ exonuclease activity of Taq DNA polymerase as well as the 3’5’ exonuclease activity of the proofreading polymerase. FideliTaq DNA Polymerase increases amplification fidelity up to 6 times over Taq DNA Polymerase alone and allows for amplification of longer product sizes(1-4). The enzyme blend generates PCR 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). Although a range of PCR product sizes can be amplified (Fig. 1), optimization may be necessary for targets longer than 10 kb.

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Fig. 1. Amplification of diverse targets by FideliTaq™ DNA Polymerase. Target (source): Lane 1: Numb, 455 bp (10 ng human genomic DNA); Lane 2: NRAGE, 1.55 kb (10 ng human genomic DNA); Lane 3: Numb, 4.6 kb (10 ng human genomic DNA); Lane 4: Lambda target, 20.7 kb (10 ng lambda DNA). A total of 25 cycles was used for lambda target and 35 cycles for human genomic DNA targets. A final concentration of 1.5mM MgCl2 was used in all reactions.

Properties:
Activator: Mg2+

Purity:
Free of detectable non-specific nucleases.

Storage Buffer:
20mM Tris-HCl, pH 8.5, 1mM DTT, 0.1mM EDTA, 100mM KCI, 50% glycerol, stabilizers.

Unit Definition:
One unit incorporates 10 nmol of total nucleotides into acid-insoluble material in 30 min at
74°C in a total volume of 50 µl(6-7).

Concentration:
5 units/µl

Assay Conditions:
The reaction mixture contains 25mM TAPS, pH 9.3 (at 25°C), 50mM KCl, 2mM MgCl2,
1mM b-ME, 200µM dNTPs, 250 µg/ml activated salmon sperm DNA and FideliTaq DNA Polymerase. Following incubation at 74°C for 10 min, acid-insoluble material is determined.

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

Functionally Tested 10X PCR Reaction Buffer (included, PN 71165 ):
100mM Tris-HCl (pH 8.6), 500mM KCl, 15mM MgCl2

Functionally Tested MgCl2 (included, PN 71167 ):
25mM Solution

References:

  1. BARNES, W. M. (1994) Proc. Natl. Acad. Sci. USA 91, 2216-2220.
  2. CHENG, S., FOCKLER, C., BARNES, W. M., AND HIGUCHI, R. (1994) Proc. Natl. Acad. Sci. USA 91, 5695-5699.
  3. BARNES, W. M. (1992) Gene 112, 29-35.
  4. CLINE, J., BRAMAN, J. C., AND HOGREFE, H. H. (1996) Nucleic Acids Res. 24, 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, 700-709.
  6. INNIS, M. A., MYAMBO, K. B., GELFAND, D. H. AND BROW, M. A. (1988) Proc. Natl. Acad. Sci. 85, 9436-9440.
  7. LAWYER, F. C., STOFFEL, S., SAIKI, R. K., MYAMBO, K., DRUMMOND, R. AND GELFAND, D. H. (1989) J. Biol. Chem. 264, 6427-6437.

Storage:
Shipped on dry ice. Store at -20°C.

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