Intact Genomics Hot start Taq is a thermostable DNA polymerase that possesses a 5´→3´ polymerase activity (1, 2) and a 5´ flap endonuclease activity (3, 4). Hot Start Taq DNA Polymerase is chemically modified that leads to complete inactivation of the polymerase until the initial heat activation step at the start of PCR. Hot start PCR reduces non-specific amplification during setup stages of the reaction and helps increase PCR specificity and sensitivity. This product is supplied with the unique Intact Genomics 10x PCR reaction buffer, containing MgCl2, which produces a final Mg2+ concentration of 1.5 mM, and 5X Magic Enhancer that enables efficient amplification of GC rich templates up to 84%.
The physical purity of this enzyme is ≥98% as assessed by SDS-PAGE with Coomassie® blue staining (see figure below).
E. coli strain expressing a Taq DNA Polymerase gene from Thermus aquaticus YT-1.
Hot Start Taq Polymerase Comparison Data:
We compare our hot start Taq Polymerase with leading competitor side by side and our enzyme is better than the competitor. A typical gel picture is shown below:
- Routine PCR
- Primer extension
- Colony PCR
- Efficient for amplifying high GC template DNA with Magic Enhancer
- Product Includes
Hot Start Taq DNA Polymerase
- 10x PCR Buffer with Mg2+
- 5x Magic Enhancer
- 10 mM dNTP (Cat. # 3291d, 3292d only)
50 mM Tris-HCl, 50 mM KCl, 1 mM DTT, 0.1 mM EDTA, 50% Glycerol, pH 7.5 @ 25 ºC
10x PCR Buffer with Mg2+
100 mM Tris-HCl pH 9.0, 15 mM MgCl2, 100 mM KCl, 80 mM (NH4)2SO4, 0.5% Igepal CA 630
One unit is defined as the amount of enzyme that incorporates 10 nmoles of dNTP into acid-insoluble form in 30 minutes at 72 ºC.
- Thaw 10x PCR Buffer, dNTP mix, primer solutions, 5X Magic Enhancer (if required) and mix thoroughly before use.
- Prepare a reaction mix according to the following table: The reaction mix typically contains all the components needed for PCR except the template DNA.
- Mix the reaction mixture thoroughly.
- Add template DNA to the individual PCR tubes containing the reaction mixture.
- Program the thermal cycler according to the manufacturer’s instructions.
Note: PCR program must start with an initial heat-activation step at 95 °C for 15 min.
A typical PCR cycling program is outlined in the following table.
- Place the PCR tubes in the thermal cycler and start the cycling program.
- EChien, A., Edgar, D.B. and Trela, J.M. (1976). J. Bact. 127, 1550-1557.
- Lawyer, F.C. et al. (1993). PCR Methods and Appl. 2, 275-287.
- Longley, M.J., Bennett, S.E. and Mosbaugh D.W. (1990). Nucleic Acids Res. 18, 7317-7322.
- Lyamichev, V., Brow, M.A. and Dahlberg, J.E. (1993). Science. 260, 778-783.