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PREMIUM Bullseye Taq DNA Polymerases

Item #:ASPCRREAG2 
Bullseye Taq DNA Polymera

Store at -20°CFor 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.

PNPCRREAG

Magnesium
Mg Free
Type
Standard
Dye
No dye
Pkg
Indiv
Enzymes
Premium
SKU Number Vol Magnesium Dye Enzymes Pkg Type Web Price Quantity Add to Cart
BE110203 500 Units Mg Free No dye Premium Indiv Standard
113.08
Each
BE110204 1000 units Mg Free No dye Premium Indiv Standard
176.08
Each
BE110208 10,000 units Mg Free No dye Premium Indiv Standard
1268.85
Each
Bullseye Taq DNA Polymera
PREMIUM Bullseye Taq DNA Polymerases
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