PREMIUM Bullseye HS-Taq DNA Polymerase

HS Taq Polymerase

5 units/µl

Store at -20°C. For in-vitro laboratory use only

General Description

Bullseye HS Taq DNA Polymerase is a modified form of Bullseye Taq DNA Polymerase, which is activated by heat treatment. A chemical moiety is attached to the enzyme at the active site, which renders the enzyme inactive at room temperature. Thus, during setup and the first ramp of thermal cycling, the enzyme is not active and misprimed primers are not extended. The result is higher specificity and greater yields when compared to standard DNA polymerases.

Once the reaction reaches optimal activating temperature, the chemical moiety is cleaved during a 15 minute heat activation step, releasing the active HS Taq DNA Polymerase into the reaction.

10X HS Buffer I

Tris-HCl, pH 8.5 (NH4)2S04, 15 mM MgCl2, 1% Tween 20.

10X HS Buffer II

An optimized buffer with a balanced ammonium/potassium concentration. May improve results with more complicated PCR reactions such as multiplex PCR.

Tris-HCl pH 8.7, Balanced KCl/(NH4)2S04, 15 mM MgCl2, 1% Tween 20.

HS Taq Storage Buffer

Enzyme is supplied in 20 mM Tris-HCl pH 8.3, 100 mM KCl, 0.1 mM EDTA, 1 mM DTT, 0.5% Tween 20®, 0.5% NP40, 50% glycerol.

Unit Definition

One unit is defined as the amount that incorporates 10 nmoles of dNTPs into acid-precipitable form in 30 minutes at 72°C under standard assay conditions.

Quality Control

Endonuclease, exonuclease and priming activities are not detected after 3 hours incubation of 1 mg of pUC19 plasmid DNA and 0.5 µg EcoR I digested lambda phage DNA at 72°C in the presence of 40 units of TEMPase DNA Polymerase.

Suggested Protocol using HS Taq DNA Polymerase

This protocol serves as a guideline. Optimal reaction conditions must be individually determined.

• 15 mM MgCl2 is present in the 10X HS Buffer I and II. However, in some applications, more than 1.5mM MgCl2 is needed. For this reason, 25mM MgCl2 is included with the kit. Table 2 provides the volume of 25mM MgCl2 to add to the master mix if a higher MgCl2 concentration is required.

  1. Thaw 10X HS Buffer I or/and 10X HS Buffer II, dNTP mix, primer solutions. It is important to mix the solutions completely before use to avoid localized concentrations of salts.

  2. Prepare a master mix according to Table 1. The master mix typically contains all the components needed for extension except the template DNA.

Table 1. Reaction components (master mix & template DNA)

Table 2. MgCl2 concentration in a 50 µL reaction