Intact Genomics Taq DNA Polymerase 2x Premix with Dye is a thermostable DNA polymerase that possesses a 5´→3´ polymerase activity (1, 2) and a 5´ flap endonuclease activity (3, 4). Ideal for primary extension reaction with DNA fragments having dA overhang on 3’ ends.
Intact Genomics Taq DNA Polymerase 2x Premix with Dye is ready to use, containing Taq DNA Polymerase, dNTPs, MgCl2, and stabilizers. It has been optimized for routine PCR applications.
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.
- Primer extension
- Colony PCR
- Taq DNA Polymerase 2x Premix with Dye
- 5x Magic Enhancer
1x Premix Composition
10 mM Tris-HCl pH 9.0 , 50 mM KCl, 1.5 mM MgCl2, 0.2 mM dNTP, 5% Glycerol, 0.08% Igepal CA 630, 0.05% TWEEN-20, 25 Units/ml Taq DNA Polymerase
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 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.
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.