SuperHot Master Mix is an optimized ready-to-use PCR mixture of Taq DNA Polymerase, antibodies to Taq DNA Polymerase, PCR buffer, MgCl 2 and dNTPs. 2x PCR Master Mix contains all components for PCR, except DNA template and primers.

Applications:

• PCR
• Primer extension
• Multiplex PCR
• Low-copy targets PCR
• Real Time PCR

Taq Master Mix Composition:

• Taq DNA Polymerase in reaction buffer: 0.1 u/µl
• Antibodies to Taq DNA Polymerase, concentration adjusted for the effective inhibition of DNA polymerase activity at 37°C
• 32 mM (NH 4 ) 2 SO 4
• 130 mM TrisHCl, pH 8.8 at 25°C
• 0.02% Tween-20
• 3 mM MgCl 2 (or 7,0 mM MgCl 2 for #119104 and #119108)
• dNTPs (dATP, dCTP, dGTP, dTTP): 0.4 mM of each

Protocol

Due to the inhibition of polymerase activity at room temperature by Anti Taq DNA Polymerase antibodies all reactions may be settled-up at room temperature. This will not result in increase of unspecific product or primer-dimers formation.

Add in a thin walled PCR tube:

• Gently vortex the sample and briefly centrifuge to collect all drops to the bottom of the tube.
• Overlay the sample with mineral oil or add an appropriate amount of wax if the thermal cycler is not equipped with a heated lid.
• Place the samples in a thermocycler and start the optimal PCR program.

Performance and purity tests: Tested for the absence of endodeoxyribonucleases and exodeoxyribonucleases. The 2x SuperHot Master Mix is tested for the amplification of a single-copy gene of mouse genomic DNA.

Endodeoxyribonuclease Assay: No detectable conversion of covalently closed circular DNA to nicked DNA was observed after incubation of 25 µl of 2x SuperHot Master Mix with 1 µg of pUC19 DNA in 50 µl for 4 hours neither at 37°C nor at 70°C.

Associated activities: Endonuclease and exonuclease activities were not detectable after 2 hours incubation of the mixture with 0.22 mg of EcoR I digested lambda DNA at 72 °C in the presence of 15 – 20 units of enzyme in SuperHot Master Mix.