2×HotStart Taq PCR Master Mix with Dye

ABP HotStart Taq PCR Master Mix is a 2× concentrated solution of HotStart Taq DNA Polymerase, dNTPs, and all of the components required for PCR, except DNA template and primers. HotStart Taq PCR Master Mix provides robust and reliable performance in PCR amplification. This pre-mixed formulation saves time and reduces contamination due to a reduced number of pipetting steps required for PCR set up. HotStart Taq PCR Master Mix contains inert, non-toxic dyes to visualize PCR mixing step, and also allows direct loading of PCR products on to gels for electrophoresis.

The HotStart Taq DNA Polymerase is a recombinant Taq DNA polymerase complexed with a proprietary antibody that blocks polymerase activity at ambient temperatures, whose enzyme activities can be re-activated after 1-3 minutes of incubation at 95°C. The HotStart Taq DNA Polymerase uses amplification conditions for regular Taq DNA Polymerase, except no polymerase activity will be present before the onset of thermal cycling. This prevents nonspecific DNA amplification and primer dimer formation.

SPECIAL FEATURES

• Convenient, ready-to-use mix.
• Hot start.
• Generates PCR products with 3’-dA overhangs.
• Incorporates modified nucleotides (e.g., biotin-, digoxigenin-, fluorescently-labeled nucleotides).

APPLICATIONS

• Routine PCR.
• High sensitivity PCR.
• High throughput PCR.
• Hot Start PCR.

Composition of HotStart Taq PCR Master Mix (2X)

0.05 U/µL HotStart Taq DNA polymerase, reaction buffer, 4 mM MgCl₂, 0.4 mM of each dNTP (dATP, dCTP, dGTP and dTTP).

General Protocol

1.  Thaw template DNA and all reagents on ice. Mix each solution by vortexing, and centrifuge briefly to collect residual liquid from the sides of the tubes.

2.   Prepare the following reaction mixture in a PCR tube on ice:
      

3.  Mix, and then briefly centrifuge the contents.

4.  Perform PCR using the recommended thermal cycling conditions outlined below:
     

5.   Analyze the amplification products by agarose gel electrophoresis.