Thermostable DNA Polymerase
Thermus aquaticus, recombinant, E. coli
| Cat. No. | Amount | Price (EUR) | Buy / Note |
|---|---|---|---|
| PCR-217S | 200 units | 65,80 | Add to Basket/Quote Add to Notepad |
| PCR-217L | 5 x 200 units | 263,30 | Add to Basket/Quote Add to Notepad |
For general laboratory use.
Unit Definition: One unit is defined as the amount of the enzyme required to catalyze the incorporation of 10 nmoles of dNTP's into an acid-insoluble form in 30 minutes at 74 °C.
Shipping: shipped on gel packs
Storage Conditions: store at -20 °C
avoid freeze/thaw cycles
Shelf Life: 12 months
Form: liquid
Concentration: 5 units/μl
Description:
KlenTaq is a truncated version of Taq DNA Polymerase, lacking its first 280 amino acids. The enzyme catalyzes the polymerization of nucleotides into duplex DNA in 5’→3’ direction in the presence of magnesium but lacks the 5’→3’ exonuclease activity of Taq polymerase. The enzyme is purified by an additional separation process to reduce contaminating bacterial DNA sequences.
In addition to routine PCR, KlenTaq is recommended for genotyping and primer extension. Compared to Taq the enzyme shows an improved fidelity and thermostability.
Content:
KlenTaq (red cap)
5 units/μl KlenTaq DNA Polymerase in 20 mM Tris-HCl, 100 mM KCl, 0.1 mM EDTA, 1 mM DTT, 0.5 % Tween-20, 0.5 % Nonidet P-40, 50 % (v/v) Glycerol, pH 8.0 (25°C)
KlenTaq Buffer (green cap), 10x conc.
200 mM Tris-HCl, 400 mM KCl, 25 mM MgCl2, pH 8.5 (25 °C)
| component | PCR-217S | PCR-217L |
| KlenTaq Polymerase | 200 units / 40 μl | 5 x 200 units / 5 x 40 μl |
| KlenTaq Buffer, 10x conc. | 400 μl | 5 x 400 μl |
PCR Reaction Set-Up:
The PCR procedure below shows appropriate volumes for a single 50 μl reaction. For multiple reactions, prepare a master mix of all components and dispense appropriate volumes into each PCR reaction tube prior to adding template DNA and primers.
Thaw up and briefly centrifuge each component before use.
Add the following components to a PCR tube:
Prepare PCR Master Mix
| comp. | cap | final conc. | 1 assay @ 50 μl |
| PCR-grade Water | white | fill up to 50 μl | |
| 10x reaction Buffer | green | 1x | 5 μl |
| dNTP Mix 10 mM | white | 200 μM each | 1 μl |
| KlenTaq | red | 2.5 units/reaction | 0.5 μl |
| forward primer (10 μM) | 0.1 - 0.5 μM | 0.5 - 2.5 μl | |
| reverse primer (10 μM) | 0.1 - 0.5 μM | 0.5 - 2.5 μl | |
| template DNA | 10 pg - 1 μg1) |
Mix and briefly centrifuge the components
Optimisation of MgCl2 concentration:
The final assay contains 3.5 mM Mg2+ as recommended for most applications. For an individual optimisation Mg2+ stock solution (#PCR-266) mybe added.
Incubate reactions in a thermal cycler
Recommended cycling conditions:
| initial denaturation | 96 °C | 2 min | 1x |
| denaturation annealing2) elongation3) | 96 °C 50 - 68 °C 72 °C | 15 - 30 sec 15 - 30 sec 30 sec - 4 min | 25 - 35x |
| final elongation (optional) | 72 °C | 2 min | 1x |
| hold | 4 - 8 °C |
For optimal specificity and amplification an individual optimisation of the recommended parameters may be necessary for each new template DNA and/or primer pair.
Related products: Mg2+ Stock, #PCR-266
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