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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:

Gel Cutting Tips, 1.1 x 6.5mm

• Safely excise bands from gels
• Avoid cross contamination
• One-handed operation for quick and accurate gel cutting
• Safer than using razor blades and won't scratch transilluminators

• 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

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°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.

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

Table 2. MgCl2 concentration in a 50 mL reaction

Table 2 provides the volume of 25 mM MgCl2 to add to the master mix if a certain MgCl2 concentration is required.

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)

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.