Definição de unidade: One unit is defined as the amount of the enzyme required to catalyze the incorporation of 10 nmoles of dNTPs into an acidinsoluble form in 30 minutes at 70 °C. 

Envio: Shipped on blue ice

Condições de armazenamento: Store at -20 °C

Avoid freeze/thaw cycles

Validade: 36 months

Concentração: 5 units/μL

 

5 units/µl Taq High Fidelity Pol in Tris-HCl, KCl, EDTA, DTT, 50% (v/v) Glycerol, pH 8.0 (25°C) and stabilizers.

 

Tris-HCl pH 9,0 (25°C), KCl, 15 mM MgCl₂.

 

Taq High Fidelity Pol is based on a blend of Taq DNA polymerase and a proofreading enzyme specially designed for highly accurate and efficient amplification. It shows excellent results with extremely long (up to 25 kb), GC-rich or other difficult templates. The enzyme blend includes a highly processive 5’→3’ DNA polymerase and possesses a 5’3’ polymerizationdependent exonuclease replacement activity. Its inherent 3’→5’ exonuclease proofreading activity results in a greatly increased fidelity of DNA synthesis compared to Taq polymerase. The enzyme is highly purified and free of bacterial DNA.

 

Taq High Fidelity Pol is characterized by a 2-fold higher fidelity compared to Taq polymerase.

 

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:

 

Note: Consider the volumes for all components listed next steps to determine the correct amount of water required to reach your final reaction volume.

Mix and briefly centrifuge the components.

 

Component	50 μl rxn	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.

 

Recommended cycling conditions:

**The annealing temperature depends on the melting temperature of the primers used

***The elongation time depends on the length of the fragments to be amplified. A time of 1 min/kb is recommended.

Note: For large fragments it is recommended in the elongation step to use 68 ºC and 1,5 - 2 min/kbp.

For optimal specificity and amplification an individual optimization of the recommended parameters may be necessary for each new template DNA and/or primer pair.