Intact Genomics Taq DNA Polymerase with Dye is a thermostable DNA polymerase that possesses a 5´→3´ polymerase activity (1, 2) and a 5´ flap endonuclease activity (3, 4). This product is supplied with 10x PCR reaction buffer, containing MgCl2, which produces a final Mg2+ concentration of 1.5 mM. Ideal for primary extension reaction with DNA fragments having dA overhang on 3’ ends.
The physical purity of this enzyme is ≥98% as assessed by SDS-PAGE with Coomassie® blue staining (see figure below).
- coli strain expressing a Taq DNA Polymerase gene from Thermus aquaticus YT-1.
Taq DNA Polymerase with Dye Comparison Data
- Routing PCR cloning
- Primer extension
- Colony PCR
- Elongation efficiency 1.0-1.2 kb/min.
- Formulated for amplifying long target DNA.
- Efficient for amplifying high GC content DNA with Intact Genomics magic enhancer
- Taq DNA Polymerase with Dye
- 10x PCR Buffer with Mg2+
- 5x Magic Enhancer
- 10 mM dNTP (Cat. # 3242d, 3242d-04 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 8.0, 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 PCR buffer, dNTP, 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.
3. Mix the reaction mixture thoroughly.
4. Add template DNA to the individual PCR tubes containing the reaction mixture.
5. Program the thermal cycler according to the manufacturer’s instructions.
A typical PCR cycling program is outlined in the following table.
6. Place the PCR tubes in the thermal cycler and start the cycling program.
1. EChien, A., Edgar, D.B. and Trela, J.M. (1976). J. Bact. 127, 1550-1557.
2. Lawyer, F.C. et al. (1993). PCR Methods and Appl. 2, 275-287.
3. Longley, M.J., Bennett, S.E. and Mosbaugh D.W. (1990). Nucleic Acids Res. 18, 7317-7322.
4. Lyamichev, V., Brow, M.A. and Dahlberg, J.E. (1993). Science. 260, 778-783.