Taq DNA Polymerase
Enzyme is purified from an E. coli strain carrying a plasmid with Taq DNA polymerase gene from Thermus aquaticus YT-1.
Reagents supplied: 10x Taq DNA polymerase Buffer with or w/o MgCl2 and 25mM MgCl2
Unit definition: One unit is defined as the amount of enzyme required to catalyze the incorporation of 10 nmoles of dNTPs into acid insoluble material in 30 minutes at 72ºC.
(SKUs: PD 001S-W, PD 001S-W/O, PD 001M-W, PD 001M-W/O, PD 001L-W, PD 001L-W/O)
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Product Details
Enzyme is purified from an E. coli strain carrying a plasmid with Taq DNA polymerase gene from Thermus aquaticus YT-1.
Reagents supplied: 10x Taq DNA polymerase Buffer with or w/o MgCl2 and 25mM MgCl2
Unit definition: One unit is defined as the amount of enzyme required to catalyze the incorporation of 10 nmoles of dNTPs into acid insoluble material in 30 minutes at 72ºC.
Taq DNA Polymerase is a thermostable enzyme that catalyzes 5’→3′ synthesis of DNA. The enzyme has no detectable 3’→5′ proofreading exonuclease activity, but possesses low 5’→3′ exonuclease activity.
PCR Guidelines:
PCR recommended reaction:
Components | 20μl assay | 50μl assay | Final Concentration/Quantity |
10x Taq pol. buffer | 2μl | 5μl | 1x |
dNTP mix/10mM each | 0.4μl | 1μl | 0.2mM each |
25μΜ forward primer | 0.4μl | 1μl | 0.5μM |
25μΜ reverse primer | 0.4μl | 1μl | 0.5μM |
Template DNA | Variable | Variable | 1-500ng |
Taq DNA pol. (5u/μl) | 0.1-0.2μl | 0.25-0.5μl | 1.25-2.5u / 50μl reaction |
Sterile ultrapure water | Up to 20 μl | Up to 50 μl |
PCR recommended conditions:
Step | Temperature | Time |
Initial denaturation | 94oC | 2 min |
25-35 cycles | ||
Denaturation | 94oC | 45 sec |
Annealing | 45-68oC* | 30 sec |
Extension | 72oC | 1min/kb |
Final extension | 72oC | 10 min |
Hold | 4oC | Indefinitely |
*Annealing temperature depends on primer Tm
10x Taq pol. buffer: 500 mM KCl, 100 mM Tris-HCl pH 8.5 @ 25oC, 15 mM MgCl2, 1% Triton X-100
Storage conditions:
- 100 mM NaCl
- 50 mM Tris-HCl (pH 8.0 @ 25oC)
- 1 mM DTT
- 0.1 mM EDTA
- 1% Triton X-100
- 50% glycerol
- Store at -20oC
Standard DNA Polymerase Assay Conditions (not PCR conditions): The polymerase activity is assayed in 10 mM Tris-HCl (pH 8.5), 50 mM KCl, 1.5 mM MgCl2, 0.1% Triton X-100, 200 μM each of dATP, dGTP, dCTP, dTTP (a mix of unlabeled and [3H] dTTP) and 12.5μg activated calf thymus DNA, in a final volume of 50 μl.
Functional Assay: Taq DNA Polymerase is tested for performance in the polymerase chain reaction (PCR) using 1.5 units of enzyme to amplify a 5000-bp region of Lambda DNA using 10ng as substrate. The resulting PCR product is visualized as a single band on an ethidium bromide-stained agarose gel.
Quality controls: Tested extensively for the absence of endo- and exodeoxyribonucleases. More details concerning quality controls could be found in Certificate of analysis.
Stability: Taq DNA polymerase is guaranteed to maintain stability for six months from the date of shipment when stored as directed.