Taq DNA Polymerase

Applications
Marligen’s Taq DNA polymerase can be used with a wide variety of PCR based assays including Primer Extension, Microarray Analysis, High-throughput PCR, DHPLC, and Colony PCR. The Taq DNA Polymerase has been validated with Marligen’s Signet™ Genotyping Panels formatted on the Luminex platform.

Source of Protein
A recombinant E. coli strain carrying the Taq DNA polymerase gene from the thermophilic organism Thermus Aquaticus YT-1.

Components
1. Taq DNA Polymerase is supplied in 20 mM Tris-HCl, pH 7.6, 100 mM NaCl, 1 mM DTT, 0.1 mM EDTA, 0.5% Tween-20, 0.5% Nonidet P-40, 50% Glycerol.

2. 10X PCR Buffer 1 containing 100 mM Tris-HCl, pH 8.6, 15 mM MgCl₂, 750 mM KCl.

Storage Conditions
Store at –20°C.

Concentration
5 units/µL

Quality Assurance
Purified free of contaminating endonucleases and exonucleases. In addition, enzyme purity is analyzed by SDS-PAGE.

PCR Guidelines
Taq DNA Polymerase is the original and most commonly used PCR enzyme. Taq excels at amplifying shorter (<5 kb) sequences from low-complexity template sources and produces robust yields with little or no optimization of reaction conditions.

A good practice, particularly when handling large numbers of similar samples, is to create two 2X master mixes – one containing Taq DNA Polymerase and buffer, and the other containing dNTPs, DNA, and primers. The DNA/primer mix should be mixed thoroughly and added to reaction vessels, and the polymerase/buffer mix similarly mixed and added to the DNA cocktail. Reactions can then be added to an appropriate thermal cycler for completion of the reaction.

Typical 50 µL PCR Reaction
On ice, prepare each of following master mixes, combine, and place in heated (to 94°C) thermal cycler.

2X DNA/Oligonucleotide Master Mix:
1.0 µL 10 mM dNTPs
1.0 µL 10 µM Forward Primer
1.0 µL 10 µM Reverse Primer
1.0 µL 500 ng/µL genomic DNA
21 µL Nuclease free water

2X Enzyme/Buffer Master Mix:
5.0 µL 10X PCR Buffer 1
I0.2 µL 5 U/µL Taq DNA Polymerase
19.8 µL Nuclease free water

General Cycling Conditions