|◀ 25 - 36 of 36 ▶|
View:

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


Looking for other alternatives?

Visit our PR1MA Polymerase page for more options! 
 


Bullseye High Fidelity PCR Master Mix


The newly developed Bullseye HFL PCR Master Mix represents the highest fidelity for a PCR product. Bullseye's HFL PCR Master Mix has eight times higher fidelity when compared to Pfu, previously considered to be the golden standard for high fidelity PCR. This Master Mix also works well for large PCR fragments. 

The Bullseye HFL PCR Master Mix is in a 2X format and contains modified, high fidelity thermal stable DNA polymerases, in a pre-optimized PCR buffer. The amplified DNA will be blunt-ended. When using this master mix for most PCR experiments, only template, primers and H2O will be needed.

SPECIAL FEATURES & APPLICATIONS
  • High fidelity with robustness
  • Large fragment PCR
  • 2X master mix - makes PCR easy and consistent
  • For cloning, promoter study, RACE, DNA sequencing and more
Item#:
BE135HFL
Your Price:
85.92
Each
This item is discontinued

PR1MA™ Taq DNA Polymerase 1X Master Mix

 

PR1MA™ DNA polymerase 1X Master Mix saves time and cost by enabling direct PCR amplification of unpurified templates. It contains a recombinant, truncated (lacks 5’ to 3’ exonuclease activity), highly thermostable DNA polymerase from the thermophilic bacterium Thermus aquaticus. The enzyme is thermostable up to 98 °C for polymerase chain reaction assays and is provided as a complete reaction master mix consisting of reaction buffer, dNTPs, MgCl2, and loading dye and only requires the addition of primers and DNA template. Once PCR is complete, the reaction products can be loaded directly into an agarose gel for analysis.

  • Lacks exonuclease activity
  • Thermotolerant up to 98 °C
  • Inhibitor Resistant
  • Ideal for colony PCR, genotyping, and GC-rich templates
  • Storage temperature: -20 °C 

Important note: Please be sure to use the buffer provided with this product to ensure optimal results.

 

*These products are intended for research use only, not for therapeutic or diagnostic purposes in humans or animals. The safety and efficacy of these products in diagnostic or other clinical uses has not been established.

 

Need great (q)PCR Regents at a great price? Try PR1MA!  Click here to order.
 
 
  • Reduces formation of non-specific products
  • Improves results from multiplex reactions
  • Essential for low copy number
  • Buffer II: potassium/ammonium buffer for multiplex reactions
  • Activated at elevated temperatures
  • Gives higher specificity and greater yields than standard Taq
  • Chemical moiety is attached to the enzyme at the active site and renders it inactive at room temperature; is cleaved during a 15 min. heat activation step
  • Prevents mispriming during setup and the first ramp of thermal cycling
Item#:
ASPCRREAG9
Discontinued, Contact us for more options!


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


Looking for other alternatives?

Visit our PR1MA Polymerase page for more options!



PR DNA Polymerase, High Fidelity, 2.5 U/µL

  • Provides higher fidelity than standard Taq DNA Polymerase
  • Produces blunt-ended fragments
  • Processes <3 kb with extremely high fidelity

 

Cat. No.

Units

10X Ammonium Buffer (MgCl2
15 mM)

MgCl2
25 mM

BE211102

250

1.5 mL

1.5 mL

BE210303

500

1.5 mL

1.5 mL

BE211104

1,000

2 x 1.5 mL

2 x 1.5 mL

BE211106

2,500

4 x 1.5 mL

4 x 1.5 mL

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

Bullseye PREMIUM PR DNA Polymerase is a thermostable enzyme with proofreading ability, which can be used in primer extension reactions and other molecular biology applications. PR Polymerase exhibits both 5'-3' DNA polymerase activity and 3'-5' proofreading exonuclease activity. It is recommended for applications, which require extremely high fidelity or blunt ending.


Optimal reaction conditions are achieved by using the 10x Ammonium buffer containing MgCl2 provided with the enzyme. 25 mM MgCl2 is also included separately, in case a higher MgCl2 concentration is required for a specific reaction.


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.

10X Ammonium Reaction Buffer
Tris-HCl pH 8.5, (NH4)2SO4, 1% Tween20,
15mM MgCl2

PR Storage Buffer
50 mM Tris-HCl (pH 8.0), 50 mM NaCl, 0.1 mM EDTA, 1 mM DTT, 50% Glycerol, 0.1% NP40, 0.1% 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 PR DNA Polymerase.

Suggested Protocol using PR DNA Polymerase
This protocol serves as a guideline. Optimal reaction conditions 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, will produce satisfactory results. Table 2 provides the volume of 25 mM MgCl2 to add to the master mix if a higher MgCl2 concentration is required.

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

Component

Vol./reaction

Final Conc.

10X Ammonium Buffer

5 µL

1X

dNTP mix
(12.5 mM of each)

0.8 µL

0.2 mM of
each dNTP

Primer A

Variable

0.1-0.5 µM

Primer B

Variable

0.1-0.5 µM

PR Polymerase

1 µL

2.5 units/reaction

Distilled Water

Variable

- - - -

Template DNA

Variable

0.1-0.5 µg/reaction

Total volume

50 µL

- - - -

Table 2. MgCl2 concentration in a 50µl reaction

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

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. PR is a proofreading enzyme and requires an extension time of 1-2 min/kb. 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.
Item#:
ASPCRREAG5
Discontinued, Contact us for more options!

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

Looking for other alternatives?

Visit our PCR Reagents page for more options! 
 

 

HS Taq Polymerase

5 units/µl

 

Cat. No.

Size

Units

10X TEMPase

Buffer I

(MgCl2 15mM)

10X TEMPase

Buffer II

(MgCl2 15mM)

MgCl2

25 mM

BE220302

250

1.5 mL

1.5 mL

1.5 mL

BE220303

500

1.5 mL

1.5 mL

1.5 mL

BE220304

1,000

2 x 1.5 mL

2 x 1.5 mL

2x 1.5 mL

BE220306

2,500

4 x 1.5 mL

4 x 1.5 mL

4x 1.5 mL

Store at -20°CFor 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)

Component

Vol./reaction

Final Conc.

10X HS Buffer I or II

5 µL

1X

dNTP mix (12.5 mM each)

0.8 µL

0.2 mM each

Primer A

Variable

0.1-1.0 µM

Primer B

Variable

0.1-1.0 µM

HS Taq DNA Polymerase

1 µL

5 units

Distilled Water

Variable

- - - -

Template DNA

Variable

Variable

TOTAL volume

50 µL

- - - -

Table 2. MgCl2 concentration in a 50 µL reaction

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

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 manufacturer's instructions. Each program must start with an initial heat activation step at 95°C for 15 minutes. 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.

Item#:
ASPCRREAG6
Discontinued, Contact us for more options!

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

Looking for other alternatives?

Visit our PR1MA Polymerase page for more options! 


General Description
Bullseye offers a product series specifically developed for the amplification of GC-rich DNA sequences. The Bullseye HS DNA Polymerase combined with GC Buffer I and GC Buffer II promote excellent amplification results with targets of varying degrees of GC content. Hot Start DNA Polymerase is a modified form of Bullseye Taq DNA Polymerase. 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. Once the reaction reaches optimal activating temperature, the chemical moiety is cleaved during a 10-15 minutes heat activation step, releasing the active Hot Start DNA Polymerase into the reaction. The GC Buffers in combination with Hot Start DNA Polymerase and the heat activation step result in a high success rate in amplification of DNA targets with high GC content.

Key Features

  • Amplification of multiple DNA targets with high GC content
  • High specificity, sensitivity and product yield
  • Diminished formation of non-specific product
  • Detection of low copy number targets

    Kit Components:
    HS DNA Polymerase in Storage Buffer
    5 U/ml Hot Start DNA Polymerase, 20 mM Tris-HCl pH 8.9, 100
    mM KCl, 0.1 mM EDTA, 1 mM DTT, 0.5% TweenÒ 20,
    0.5% NP40, 50% glycerol.
    4x GC Buffer I
    Optimized buffer components, 6 mM MgCl
    4x GC Buffer II
    Optimized buffer components, 6 mM MgCl
    MgCl
    25 mM MgCl in PCR grade water

Item#:
ASPCRREAG7

Need great (q)PCR Regents at a great price? Try PR1MA!  Click here to order.



HS Taq Polymerase, 2X Mix

with HS Buffer I

 

Cat.No.

Size Reactions

HS Taq, 2X Mix (Buffer I)

Final MgCl2

Conc.

BE230301

100

2X HS Buffer I Mix

1.5mM

BE230303

500

2X HS Buffer I Mix

1.5mM

BE230304

1,000

2X HS Buffer I Mix

1.5mM

BE230306

2,500

2X HS Buffer I Mix

1.5mM

Store at -20°CFor in-vitro laboratory use only

 

General Description

Bullseye HS Taq Polymerase, 2X Mix is a ready-to-use 2.0X master mix. Simply add primers, template, and water to successfully carry out primer extensions and other molecular biology applications.

Bullseye HS Taq Polymerase Mix, the NH4+ buffer system, dNTPs and magnesium chloride are present in HS Taq Pol Mix with HS Buffer I. Each reaction requires 25 µL of the 2.0X reaction mix. Simply add primers, template and water to a total reaction volume of 50 µL.

Bullseye" HS Taq Polymerase Mix 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.

Bullseye" HS Taq 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 HS Taq Pol, 2x Mix

Tris-HCl pH 8.5, (NH4)2S04, 3.0mM MgCl2, 0.2% Tween 20Ã, 0.4 mM dNTPs, 0.2 units/µL HS Hot Start DNA Polymerase

Stabilizer

Suggested Protocol using HS Taq Pol, 2x Mix

This protocol serves as a guideline only. Optimal reaction conditions may vary and must be individually determined.

  • Set up reaction mixtures in an area separate from that used for DNA preparation or product analysis.
  • The table below shows the reaction set up for a final volume of 50 mL.
  • Important: Mix the solutions completely before use to avoid localized concentrations of salts.

    1. Set up each reaction as follows:

Component

Vol./Reaction

Final Conc.

HS Hot Start Master Mix

with HS Bufffer I

25 µL

1X

Primer A

Variable

0.11.0 µM

Primer B

Variable

0.11.0 µM

Distilled Water

Variable

- - - -

Template DNA

Variable

Variable

TOTAL volume

50 µL

- - - -

2. Mix gently by pipetting the solution up and down a few times.

3. Program the thermal cycler according to the manufacturer's instructions.

4. Each program must start with an initial heat activation step at 95°C for 15 minutes.

For maximum yield and specificity, temperatures and cycling times should be optimized for each new template target or primer pair.

A typical thermal cycling program is shown below:

95°C for 15 min. Activate HS Hot Start Polymerase

30-40 cycles:

95°C 30 sec Denature template

45-65°C 30 sec Anneal primer

72°C 1-5 min Elongation

72°C for 5 min Elongation

5. Place the tubes in the thermal cycler and start the reaction.

Item#:
ASPCRREAG8
While Supplies Last!

This brand is being discontinued and will only be available while supplies last.

 

Need great (q)PCR Regents at a great price?

Try our PR1MA™ Taq and save!

 
  • 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
Item#:
ASPCRREAG1
Discontinued, Contact us for more options!


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


Looking for other alternatives?

Visit our PR1MA Polymerase page for more options!



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&#37;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.

Item#:
ASPCRREAG2

This brand is being discontinued and will only be available while supplies last.


Looking for other alternatives?

Visit our PR1MA Polymerase page for more options!


 

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)

Item#:
ASPCRREAG3
Discontinued, Contact us for more options!

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

Looking for other alternatives?

Visit our PR1MA Polymerase page for more options! 





R-Taq DNA Polymerase 5 units/µL
 
 

Cat. No.Units10X Ammonium Buffer
(MgCl2 15mM)
MgCl2
25 mM
BE2003035001.5 mL1.5 mL
BE2003041,0002x 1.5 mL2x 1.5 mL
BE2003062,5004x 1.5 mL4x 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)

Item#:
ASPCRREAG4

PR1MA™ T4 Gene 32 Protein

 

PR1MA™ T4 Gene 32 Protein (T4 gp32) is a single-stranded DNA binding (ssDNA) protein required for E. coli bacteriophage T4 replication. It binds and stabilizes ssDNA structures which facilitates electron microscopic examination, and has also been shown to improve restriction digests, improve T4 DNA polymerase activity, and increase the yield of PCR reactions, including those with long amplicons. 

  • Optimal temperature: 37 °C
  • Heat inactivation: 65 °C for 20 minutes
  • Storage temperature: -20 °C
  • 10X T4 gp32 reaction buffer included

Buffer composition

  • 50% glycerol
  • 50 mM Tris-HCl
  • 50 mM KCl
  • 1 mM DTT
  • 0.1  mM EDTA
  • 0.1% Tween-20
  • pH = 7.5 

*These products are intended for research use only, not for therapeutic or diagnostic purposes in humans or animals. The safety and efficacy of these products in diagnostic or other clinical uses has not been established. 
 

|◀ 25 - 36 of 36 ▶|
View: