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

Applications:

• Regular PCR up to 5 kb
• PCR with bacterial DNA

Taq Master Mix Composition:

• Taq DNA Polymerase (recombinant) in reaction buffer: 0.1 units/ µl
• 32 mM (NH 4 ) 2 SO 4
• 130 mM Tris-HCl, pH 8.8 at 25°C
• 0.02% Tween-20
• 5.5 mM MgCl 2
• dNTPs (dATP, dCTP, dGTP, dTTP): 0.4 mM of each

Protocol

It’s strongly recommended to settle-up all reactions in ice to avoid formation of unspecific products 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.

If MgCl 2 is not added to the reaction mixture, final concentration of MgCl 2 in the reaction mixture will be 2.75 mM.

Addition of 1 µl of 100 mM MgCl 2 into the mixture results in concentration of MgCl 2 4.75 mM in the reaction mixture for 50 µl reaction volume.

Performance and purity tests: Tested for the absence of endodeoxyribonucleases and exodeoxyribonucleases. Performance is tested on single-copy genes of human and mouse DNA.

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