1x8 FilmStrips™ for Sealing Only One 8-well Row of a 96-well Plate

SmartSeal™ Semi-Automated Microplate Sealer

• 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:

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

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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&#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.

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.