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- Excellent all-purpose amplification enzyme
- Thermostable recombinant DNA polymerase from Thermus aquaticus
- Exhibits very high activity in primer extension
- Has both a 5' to 3' DNA polymerase and a 5' to 3' exonuclease activity
- Does not have 3' to 5' exonuclease activity-no proofreading ability
- Leaves an A-overhang, which makes the enzyme ideal for TA cloning
- Includes MgCl² in buffer
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Store at -20°C. For in-vitro laboratory use only
General Description
Bullseye Taq DNA Polymerase is a thermostable recombinant DNA polymerase, which exhibits very high activity in primer extension and other molecular biology applications. The enzyme is isolated from Thermus aquaticus and has a molecular weight of approximately 94 kDa.
Bullseye Taq DNA Polymerase has both a 5'®3' DNA polymerase and a 5'®3' exonuclease activity. The enzyme lacks a 3'®5' exonuclease activity (no proofreading ability). Taq DNA Polymerase leaves an A¢ overhang, which makes the enzyme ideal for TA cloning.
The 10X Reaction Buffer provided does not contain Mg+2. 25 mM MgCl2 is supplied separately.
10X Mg++ Free Standard Buffer
100 mM Tris-HCl pH 8.3, 500 mM KCl.
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.
Storage and Dilution 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.
Quality Control
Each lot of Taq DNA Polymerase is tested for contaminating activities, with no trace of endonuclease activity, nicking activity, exonuclease activity or priming activity.
Suggested Protocol using Taq DNA Polymerase
This protocol serves as a guideline only. Optimal reaction conditions may vary and must be individually determined.
n In some applications, MgCl2 is needed for the best results. For this reason, 25 mM MgCl2 is included with the kit.
1. Thaw 10X Mg2+ Free Standard Buffer, 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)
Component |
Vol./reaction |
Final Conc. |
10X Mg2+ Free Buffer |
5 µl |
1X |
MgCl2, 25 mM |
1- 9 µl |
0.5 4.5 mM |
dNTP mix (12.5 mM of each) |
0.8 µl |
0.2 mM of each dNTP |
Primer A |
Variable |
0.11.0 µM |
Primer B |
Variable |
0.11.0 µM |
Taq DNA Polymerase |
Variable |
1-5 units |
Template DNA |
Variable |
Variable |
Distilled Water |
Variable |
- - - - |
TOTAL volume |
50 µl |
- - - - |
Table 2. MgCl2 concentration in a 50 mL reaction
Final MgCl2 conc. in reaction (mM) |
0.5 |
1.0 |
1.5 |
2.0 |
2.5 |
3.0 |
3.5 |
4.0 |
4.5 |
Volume of 25 mM MgCl2 per rxn (µl): |
1 |
2 |
3 |
4 |
5 |
6 |
7 |
8 |
9 |
Table 2 provides the volume of 25 mM MgCl2 to add to the master mix if a certain MgCl2 concentration is required.
3. Mix the master mix thoroughly and dispense appropriate volumes into reaction tubes. Mix gently, e.g., by pipetting the master mix up and down a few times.
4. Add template DNA to the individual tubes containing the master mix.
5. Program the thermal cycler according to the manufacturers instructions. For maximum yield and specificity, temperatures and cycling times should be optimized for each new template target or primer pair.
6. Place the tubes in the thermal cycler and start the reaction.
2x Master Mix Kit (1.5 mM MgCl2)
Bullseye Taq DNA Polymerase Mix is a ready-to-use 2x reaction mix. Simply add primers, template, and water to successfully carry out primer extensions and other molecular biology applications.
Bullseye Taq polymerase, the NH4+ buffer system, dNTPs, and magnesium chloride are conveniently present in the Taq DNA Polymerase Mix. (Inert Red Dye is present in BE180303 only)
Bullseye Taq DNA Polymerase Mix offers several advantages. Set up time is significantly reduced. The chance of contaminating component stocks is eliminated. Reduction of reagent handling steps leads to better reproducibility. Standard tests can be set up with the confidence that results will be consistent every time.
Composition of 2x Taq Master Mix
150 mM Tris-HCl pH 8.5, 40 mM (NH4)2S04, 3 mM MgCl2, 0.2%
Tween 20Ò
4 mM dNTPs
2 units/µL AS ONE Taq polymerase
Inert Red Dye & Stabilizer (BE180303 only)
R-Taq DNA Polymerase 5 units/µL
Cat. No. | Units | 10X Ammonium Buffer (MgCl2 15mM) | MgCl2 25 mM |
BE200303 | 500 | 1.5 mL | 1.5 mL |
BE200304 | 1,000 | 2x 1.5 mL | 2x 1.5 mL |
BE200306 | 2,500 | 4x 1.5 mL | 4x 1.5 mL |
Store at -20°C. For in-vitro laboratory use only
General Description
Bullseye R-Taq DNA Polymerase is a thermostable recombinant DNA polymerase, which exhibits very high activity in primer extension and other molecular biology applications. R-Taq contains a red dye which provides easy and quick identification of reactions to which enzyme was added and allows confirmation of complete mixing. The inert dye has no effect on downstream processes. R-Taq is added directly to the reaction mix and is used in the same manner as standard Taq DNA Polymerase.
Bullseye R-Taq DNA Polymerase has both a 5'®3' DNA polymerase and a 5'®3' exonuclease activity. The enzyme lacks a 3'®5' exonuclease activity. R-Taq DNA Polymerase leaves an A-overhang, which makes the enzyme ideal for TA cloning.
- High performance thermostable DNA polymerase
- Red dye identifies tubes which contain enzyme and confirms complete mixing of reagents
- Leaves an A-overhang
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.
Storage Buffer
Enzyme is supplied in 20 mM Tris-HCl pH 8.3, 100 mM KCl, 0.1 mM EDTA, 1 mM DTT, inert dye, 0.5 % Tweenà 20, 0.5% NP40, 50% glycerol.
Component | Vol./reaction | Final Conc. |
10X Ammonium Buffer | 5 µL | 1X |
dNTP mix (12.5 mM each) | 0.8 µL | 0.2 mM each dNTP |
Primer A | Variable | 0.1-0.5 µM |
Primer B | Variable | 0.1-0.5 µM |
R-Taq DNA Pol | 1 mL | 5 units/reaction |
Distilled Water | Variable | - - - - |
Template DNA | Variable | 0.1-0.5 µg/reaction |
TOTAL volume | 50 µL | - - - - |
Table 2. MgCl2 concentration
3. Mix the master mix thoroughly and dispense appropriate volumes into reaction tubes. Mix gently (e.g., by pipetting) the master mix up and down a few times.
4. Add template DNA (0.1-0.5 mg/reaction) to the individual tubes containing the master mix.
5. Program the thermal cycler according to the manufacturer's instructions.
For maximum yield and specificity, temperatures and cycling times should be optimized for each new template target or primer pair.
6. Place the tubes in the thermal cycler and start the reaction.
7. After primer extension, load 5-10 mL of a 50 mL reaction directly on an agarose gel for analysis.
Tween 20 is a registered trademark of ICI Americas, Inc.
Final MgCl2 conc. in reaction (mM) | 1.5 | 2.0 | 2.5 | 3.0 | 3.5 | 4.0 | 4.5 |
Additional volume of 25 mM MgCl2 per reaction (µL): | 0 | 1 | 2 | 3 | 4 | 5 | 6 |
10X Ammonium Reaction Buffer
Tris-HCl pH 8.5, (NH4)2S04, 15 mM MgCl2, 1% Tween 20
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 R-Taq DNA Polymerase.
Suggested Protocol using R-Taq Polymerase
This protocol serves as a guideline. Optimal reaction conditions such as incubation times, temperatures, and amount of template DNA may vary and must be individually determined.
1. Thaw 10X Ammonium Buffer, dNTP mix, and 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.
The optimal MgCl2 concentration should be determined empirically but, in most cases a concentration of 1.5 mM, as provided in the 1X Ammonium Buffer, produces satisfactory results. Table 2 provides the volume of 25mM MgCl2 to add to the master mix if a higher MgCl2 concentration is required.
Table 1. Reaction components (master mix & template DNA)
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