Introduction

PCR Master Mix provides an efficient, accurate, and convenient way to amplify DNA from any template. This pre-mixed formulation saves time and reduces contamination due to a reduced number of pipetting steps required for PCR set up.

PCR Master Mixis a 2X concentrated ready-to-use solution of Taq DNA polymerase, dNTPs, MgCl2, PCR stabilizers and reaction buffers at optimal concentrations required for routine PCR DNA amplification. To set up a PCR reaction, DNA template, primers and water need to be added to the Master Mix at proportions as given in the protocol below. The PCR Master Mix is optimized to amplify targets up to 5 kb in length from lambda DNA. Additionally included gel loading reagent and dye allow PCR products to be loaded onto agarose gel directly.

The Master Mix provides a final concentration of 1.5 mM MgCl2 in the reaction mix. If a higher Mg2+ concentration is desired, please adjust Mg2+ concentration accordingly using 25 mM Mg2+ solution provided with the Master Mix in the kit.

Taq DNA Polymerase is a thermostable DNA polymerase isolated from a strain of Thermus Aquaticus, possessing a 5´→ 3´ polymerase activity and a very low 5´→ 3´ exonuclease activity. It has a half life of 3 hours at 95o C, and can replicate a 1000 base pair strand of DNA in less than 10 seconds at 72 °C. Taq has a middle level of fidelity with an error frequency of 1.10-5(around 1 mutation in 100,000 base pairs) during DNA synthesis. It is designed for use in primer extension reaction as well as sequencing. DNA sequencing with Taq DNA Polymerase produces uniform bands intensities and low background.