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Home > PCR, Protein, RNA, DNA > PCR > Reagents > PCR Reagents > PREMIUM Bullseye R-Taq DNA Polymerase

PREMIUM Bullseye R-Taq DNA Polymerase

Discontinued, Contact us for more options!

Item #: ASPCRREAG4

This brand is being discontinued, please contact us for other options.

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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 % TweenÃ 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):

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)

PNPCRREAG

Magnesium

Enzymes

Premium

Pkg

Indiv

Type

Standard

SKU Number	Vol	Magnesium	Dye	Enzymes	Pkg	Type	Price	Quantity	Add to Cart
BE200304	1,000 Units	Mg	Red Dye	Premium	Indiv	Standard	240.6200 240.62 Each		Item is no longer available.
BE210303	500 Units	Mg	No Dye	Premium	Indiv	Proof Reading	298.5300 298.53 Each		Item is no longer available.