Klentaq DNA Polymerase
Thermostable DNA Polymerase
Thermus aquaticus, recombinant, E. coli
| Catálogo Nº | Apresentação | Preço (R$) | Comprar |
|---|---|---|---|
| POL-127XS | 250 U | 249,94 | Adicionar ao Carrinho |
| POL-127S | 500 U | 465,63 | Adicionar ao Carrinho |
| POL-127M | 1.000 U | 699,30 | Adicionar ao Carrinho |
| POL-127L | 2 x 1.000 U | 1.398,60 | Adicionar ao Carrinho |
| POL-127XL | 4 x 1.000 U | 2.798,25 | Adicionar ao Carrinho |
Condições de armazenamento: Store at -20 °C
Validade: 12 months
Descrição:
Klentaq is a 5'- exonuclease deficient Taq polymerase (an Nterminal deletion of Taq) with improved fidelity (about 25%) and thermostability (about 2 degrees) relative to wild-type Taq, and it gives higher yields of amplicon.
Contente:
Kit contents:
Klentaq (blue cap)
5 units/μl Klentaq DNA Polymerase in Tris-HCl pH 8.0 (25 °C), KCl,
EDTA, DTT, 50% (v/v) Glycerol and stabilizers.
Klentaq Reaction Buffer complete (red cap) - 10x conc.
Tris-HCl pH 8.5 (25°C), KCl and 25 mM MgCl2.
PCR Reaction Setup
The PCR procedure below shows appropriate volumes for a single 50 μL reaction. For multiple reactions, prepare a master mix of components common to all and then dispense appropriate volumes into each PCR reaction tube prior to adding template DNA and primers. Thaw, mix, and briefly centrifuge each component before use. Add the following components to a microcentrifuge tube:
1. Prepare PCR master mix
Note: Consider the volumes for all components listed next steps to determine the correct amount of water required to reach your final reaction volume.
1. Prepare PCR master mix
Note: Consider the volumes for all components listed next steps to determine the correct amount of water required to reach your final reaction volume.
| Component | 50 μl assay | Final conc. |
| PCR grade water | to 50 μl | - |
| 10x Reaction Buffer Complete | 5 μl | 1x |
| dNTP Mix 10 mM | 1 μl | 200 μM |
| KlenTaq DNA Pol (5 U/μl) | 0,5 μl | 2,5 U/assay |
| template DNA | x μl | <500 ng |
Mix and briefly centrifuge the components.
2. Add template DNA and primers
| Component | 50 μl assay | final conc. |
| Forward primer (10 μM) | 0,5 – 2,5 μl | 0,1 – 0,5 μM |
| Reverse primer (10 μM) | 0,5 – 2,5 μl | 0,1 – 0,5 μM |
| Template DNA | x μl | 10 pg – 1 μg* |
*genomic DNA: 1 ng – 1 μg; plasmidial DNA: 1 pg – 1 ng
Cap each tube, mix, and briefly centrifuge the content.
4. Incubate reactions in a thermal cycler
Recommended cycling conditions:
| Initial denaturation | 95 °C | 2 min | 1x |
| Denaturation | 95 °C | 15 - 30 sec | 25 - 40 cycles |
| Annealing1 | 45 - 68 °C | 15 - 30 sec | 25 - 40 cycles |
| Elongation2 | 68 °C | 1 min/kbp | 30 cycles |
| Final extension (optional) | 68 °C | 1 – 2 min/kb | 1x |
| Hold | 4 – 8 °C | ∞ | 1x |
1)The annealing temperature depends on the melting temperature of the primers used
2)The elongation time depends on the length of the fragments to be amplified. A time of 2 min/kb is recommended.
For optimal specificity and amplification an individual optimization of the recommended parameters may be necessary for each new template DNA and/or primer pair.