Taq DNA Polymerase

UA070125: 5 U/μl Taq DNA Polymerase in 10 mM Tris-HCl, 100 mM KCl, 1 mM DTT, 0.1 mM EDTA, 0.5%Tween 20, IGEPAL® CA-630, 50%Glycerol (pH7.4@ 25°C)
10* Reaction Buffer: 100 mM Tris-HCl, 500 mM KCl, 15 mM MgCl2 (pH 8.3 @ 25°C)

a. Dissolve and mix the various solutions required for the PCR reaction. Place Taq DNA Polymerase in an ice bath or an ice box.
b. Refer to the table below to set up the PCR reaction system on the ice bath (if there are multiple similar PCR reactions, you can first prepare a large volume mixture containing water, buffer, dNTP, and Taq enzyme, and then divide it into each PCR reaction tube. According to the situation, sometimes the mixture can include primers):

Notes: Gently mix the reaction. Collect all liquid to the bottom of the tube by a quick spin if necessary. Overlay the sample with mineral oil if using a PCR machine without a heated lid.
c. Transfer PCR tubes from ice to a PCR machine with the block preheated to 95°C and begin thermocycling.
Thermocycling conditions for a routine PCR:

1. A balanced low concentration of dNTP is more favorable for enzyme activity and reduces mismatches, yielding a high amount of specific DNA reaction products;

2. Taq DNA polymerase, like many other polymerases, is a Mg2+-dependent enzyme and is very sensitive to the concentration of Mg2+. 2mM Mg2+ can meet most PCR amplification, and for some PCR, in order to ensure better amplification, it can be adjusted to 2-4 mM.