HS Taq Polymerase Master Mix BLUE is a ready to use 2 X master mix. Simply add primers, template and water to successfully carry out primer extensions. The master mix is available in 100, 500, 1000, and 2500 reaction sizes and is available with two different buffer options (HS Buffer 1 and HS Buffer 2)

HS Taq Polymerase is a modified form of Bullseye Taq, activated by heat treatment. This results in higher specificity and greater yields when compared to standard Taq. The added benefit of the HS Master Mix Blue is that it can be loaded directly onto the gel as there is no need to use separate loading dyes for subsequent electrophoresis and visualization. This cuts down on the chance of contaminating the component stocks and leads to better reproducibility.

Composition of 2 X HS Master Mix Blue with Buffer 1
Tris-HCl, pH 8.5, (NH4)2SO4,3 mM, MgCl2, .2 % Tween 20, .4 mM dNTPs, .2 units/uL HS Taq Polymerase, Inert Blue Dye, Stabilizer

Composition of 2 X HS Master Mix Blue with Buffer 2
Tris-Hcl, pH 8.5, Balanced KCl/(NH4)2SO4,3 mM, MgCl2, .2 % Tween 20, .4 mM dNTPD, .2 units/uL HS Taq Polymerase, Inert Blue Dye, Stabilizer

New and Improved! Ideal for General PCR, Genomic analysis and TA cloning!

Bulleye Taq DNA Polymerase is the most popular thermostable enzyme used in DNA amplification experiments. This high performance Taq DNA polymerase is specifically purified to produce excellent yields with little or no background. Its outstanding activity and unique thermal DNA amplification properties make it one of the best-value DNA polymerases available.

• Robust performance
• Leaves 3' A overhang
• Stable at all storage temperatures
• Ideal for general PCR, genomic analysis and TA cloning
• This product is not recommended for work with neo-primers

Storage: -20°C

Quality Control:
Taq DNA Polymerase is highly purified, free of contaminating endonucleases, exonucleases and nicking activity. For endonuclease assay, 1µg of Lamda/Hind III DNA is incubated with 20 units of enzyme in assay buffer at 75oC for 16 hours with no visible contaminating activity observed. Also, every lot is tested for its performance consistency.

Unit Definition:
One unit incorporates 10nmoles of 4 radioactive labeled dNTPs into acid-insoluble material in 30 minutes at 74°C.

Storage Buffer:
5 units/µl in 50mM Tris-HCl (pH8.0), 100mM NaCl, 0.1mM EDTA, 1mM DTT, 50% glycerol, 0.5% TritonX-100, and 0.5% NP-40.
10x Reaction Buffer 100mM KCl, 100mM Tris HCl (pH9.0), 80mM (NH4)2SO4, and 1.0% Triton X-100. (Mg++free): is optimized for use with 200µM dNTPs.

Magnesium Chloride:
25mM MgCl2: In general, 1.5mM MgCl2 is recommended; this may vary with different conditions and primer sets.

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For laboratory research only. Not for clinical applications.

• Pre-optimized ready-to-use PCR reagents
• Less pipetting to avoid contamination
• Quick and easy to set up
• Direct loading for electrophoresis
• Reproducible results

Users only need to add templates and primers, and water if needed. Extra 25mM magnesium solution is also provided for additional fine adjustments. The Red Mixes contain a non-hazardous inert red dye. The purple mixes contain a non-hazardous inert purple dye. Completed PCR reactions utilizing the D124-R (red dye) or D124-P (purple dye) can be directly loaded into a well of agarose gel or other gels, for electrophoresis. No extra loading buffer is needed.

Storage: 4°C for up to one month, or -20°C for long term storage.

Magnesium Chloride: In general, 1.5mM MgCl2 is recommended; this may vary with different conditions and primer sets. Some primers/templates may require adjustments for MgCl2 concentration, which can be achieved as shown below:

Directions for use: For a 50µl reaction, use 25µl of the Taq Master Mix, add template, primers, and water to a final volume of 50µl. Cycling conditions vary for different templates and primers. To start with, try 30 cycles as follows: denature at 94°C for 30 seconds, anneal around 55°C for 30 seconds, and extend at 72°C for 1 minutes/kb. After the PCR cycles, extend at 72°C for another five minutes to complete the PCR. Then store the reaction at 4°C.

1x Composition: 10mM KCl, 20mM Tris HCl (pH9.0), 16mM (NH4)2SO4, 0.1% Triton X-100, 1.5mM MgCl2, 200mM dNTPs, 2.5units/25ul of Taq DNA polymerase, (optional: trace amount of red or purple dye) and enzyme stabilizers.

Bullseye GC HS 2x Master Mix I is an all-in-one 2x master mix containing HS DNA polymerase, GC Buffer I, enhancer, dNTPs and MgCl2. Simply mix GC HS 2x Master Mix I with primers, template and water and you are ready to carry out successful primer extensions. HS 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, increased sensitivity and greater yields when compared to standard DNA polymerases.

Key Features

• For amplification of DNA targets with high GC content
  
• Convenient reaction set-up at room temperature
  
• High specificity, sensitivity and product yield
  
• Detection of low abundance targets
  
• Diminished formation of non-specific product

   

  Composition of GC HS 2x Master Mix I
  

• HS DNA Polymerase
  
• Optimized buffer components, 3.0 mM MgCl
  
• dNTPs
  
• Enhancer

General Description
Bullseye GC HS 2x Master Mix II is an all-in-one 2x master mix containing HS DNA polymerase, GC Buffer II, enhancer, dNTPs and MgCl. Simply mix GC HS 2x Master Mix II with primers,template and water and you are ready to carry out successful primer extensions. HS 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, increased sensitivity and greater yields when compared to standard DNA polymerases.

Key Features

• For amplification of DNA targets with high GC content
  
• Convenient reaction set-up at room temperature
  
• High specificity, sensitivity and product yield
  
• Detection of low abundance targets
  
• Diminished formation of non-specific product

  
  Composition of GC HS 2x Master Mix I
  

• HS DNA Polymerase
  
• Optimized buffer components, 3.0 mM MgCl
  
• dNTPs
  
• Enhancer

  
  Storage and Stability
  The unopened kit is stable at -20°C for 1 year

Combo Includes:
200 μL Taq DNA Polymerase (5 units/μL)
4 x 500 μL vials of dNTP Premix (100 mM each of dATP, dCTP, dGTP, dTTP)

D118-1000

High-performance Taq DNA Polymerase specially purified to produce excellent yields with little or no background. Its outstanding activity and unique thermal DNA amplification properties make it one of the most economic DNA polymerases available.

Special Features

• Robust performance
• Leaves 3'A overhang
• Stable at all storage temperatures
• Best price/quality
• APPLICATIONS
• General PCR
• Genomic analysis
• TA cloning

D116

Ready-to-use, ultra-pure, molecular grade solution made from 100mM each of dATP, dCTP, dGTP, and dTTP stock. The PreMix is designed to reduce "hands-on" time for researchers, thereby lowering the possibility of contamination and error. Furthermore, every lot of dNTP solution is pre-tested for optimal performance. The nucleotides are supplied in sterile, doubly distilled water. Each dNTP has a final concentration of 10mM for a total PreMix dNTP concentration of 40mM.

Special Features

• Ready to use
• Each lot functionally tested
• Purity; at least >99%
• APPLICATIONS
• For RT-PCR, large PCR, and real-time PCR
• Reverse Transcription and mutagenesis

• 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

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

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)

Table 2. MgCl2 concentration in a 50µl reaction

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.

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

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

HS Taq Polymerase, 2X Mix

with HS Buffer I

Store at -20°C. For 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: