Top DNA Polymerase


Enzymes for everyday PCR, faster than Taq DNA Polymerase, and TA Cloning compatible

Top DNA polymerase is a novel thermostable DNA polymerase that is more processive than Taq DNA polymerase. In fact, the extension rate of Top DNA Polymerase is > 3 x that of Taq DNA Polymerase! Top DNA Polymerase can be used for a variety of PCR applications (including TA cloning) and is a robust enzyme for everyday PCR. It contains no proofreading or 5’-3’ Exonuclease activity.
 

Features and Benefits

  •  Fast: More than three times more processive than standard Taq DNA Polymerase
  •  High performance: Amplifies fragments up to 10 kb
  •  Value:  No license fee to pay!
     

]]> Enzyme Properties

5' to 3' exonuclease activity: No
3' to 5' exonuclease activity: No
3’ – A Overhang: Yes
Fragment Size: Up to 10 kb

 

Application

Standard PCR, primer extension, TA cloning and gene sequencing

 

Reagents Supplied

10 x Reaction Buffer with (or without) MgCl2: Tris (pH 9.0), 15 mM MgCl2, etc
1 x Dilution Buffer: 50% glycerol containing 20 mM Tris-HCl, 0.5 mM EDTA, 1 mM DTT, 100 mM KCl, stabilizers, pH 8.0
dNTP Mix: 2.5 mM of each dNTP

 

Concentration

5 U/μl

 

Storage Conditions

50% glycerol containing 20 mM Tris-HCl, 0.5 mM EDTA, 1 mM DTT, 100 mM KCl, stabilizers, pH 8.0

 

Store Temperature

-20°C

 

Unit Definition

One unit is defined at the amount of enzyme that will incorporate 10 nmole of dNTP into acid-insoluble material in 30 minutes at 72°C.



Figure 1. Sensitivity test of Top DNA polymerase and Taq DNA polymerase using Lambda genomic DNA.
Each fragment was amplified from a template dilution series (100 ng to 10 fg DNA per reaction) using 1 U of each DNA Polymerase.

Lane MW : 1 kb DNA Ladder (D-1040)
Lane 1: 100 ng Lambda genomic DNA
Lane 2: 10 ng Lambda genomic DNA
Lane 3: 1 ng Lambda genomic DNA
Lane 4: 100 pg Lambda genomic DNA
Lane 5: 10 pg Lambda genomic DNA
Lane 6: 1 pg Lambda genomic DNA
Lane 7: 100 fg Lambda genomic DNA
Lane 8: 10 fg Lambda genomic DNA



Figure 2. Sensitivity test of Top DNA Polymerase and Taq DNA polymerase using bacterial and human genomic DNA.
A 500 bp fragment was amplified from a bacterial genomic DNA dilution series (Lane 1-4: 1 ng to 1 pg per reaction) and a 220 bp fragment was amplified from a human genomic DNA dilution series (Lane 5-8: 10 ng to 10 pg per reaction).
1 U of each DNA Polymerase was used for all reactions.

Lane MW : 100 bp plus DNA Ladder (D-1035)
Lane 1: 1 ng bacterial genomic DNA
Lane 2: 100 pg bacterial genomic DNA
Lane 3: 1 ng Lambda genomic DNA
Lane 4: 1 pg bacterial genomic DNA
Lane 5: 10 ng human genomic DNA
Lane 6: 1 ng human genomic DNA
Lane 7: 100 pg human genomic DNA
Lane 8: 10 pg human genomic DNA



Figure 3. Enzyme activity test of Top DNA Polymerase and Taq DNA polymerase.
Top DNA polymerase/ Taq DNA polymerase was serially diluted and used to amplify 20 ng of each Lambda and human genomic DNA.

Lane MW : 100 bp plus DNA Ladder (D-1035)
Lane 1: 1 U of Top DNA Polymerase used
Lane 2: 0.5 U of Top DNA Polymerase used
Lane 3: 0.33 U of Top DNA Polymerase used
Lane 4: 0.25 U of Top DNA Polymerase used
Lane 5: 1 U of Taq DNA Polymerase used
Lane 6: 0.5 U of Taq DNA Polymerase used
Lane 7: 0.33 U of Taq DNA Polymerase used
Lane 8: 0.25 U of Taq DNA Polymerase used



Figure 4. Long PCR amplification test of Top DNA Polymerase and Taq DNA Polymerase using Lambda DNA.
10 ng of Lambda DNA and 1 U of each DNA Polymerase used for amplification.

M1: 1 kb DNA Ladder (D-1040)
M2: Lambda DNA/Hind lll Marker (D-1050)
Lane 1: 2 kb PCR product
Lane 2: 2 kb PCR product
Lane 3: 2 kb PCR product
Lane 4: 2 kb PCR product
Lane 5: 2 kb PCR product
Lane 6: 2 kb PCR product
Lane 7: 2 kb PCR product
Lane 8: 2 kb PCR product
Lane 9: 2 kb PCR product



Figure 5. Sensitivity test using Real-time qPCR with SYBR Green and Top DNA Polymerase.
The standard curve shows a high correlation of R2 = 0.9993.

Lane MW : 100 bp plus DNA Ladder (D-1035)
Lane 1: 1 ng bacterial genomic DNA
Lane 2: 100 pg bacterial genomic DNA
Lane 3: 1 ng Lambda genomic DNA
Lane 4: 1 pg bacterial genomic DNA
Lane 5: 10 ng human genomic DNA
Lane 6: 1 ng human genomic DNA
Lane 7: 100 pg human genomic DNA
Lane 8: 10 pg human genomic DNA


 






 

]]> Manual
• Top DNA Polymerase (E3100)

• Top DNA Polymerase (E3100-1)

• Top DNA Polymerase (E3100-2)

• Top DNA Polymerase (E3100-3)


MSDS
• D-3001

• Top DNA Polymerase

• Top DP-dilution buffer

• Top DP-reaction buffer

• 20mM MgCl2


Brochure
• Enzymes 2010 Brochure


Quality Assurance
Bioneer is the holder of Quality Management System Certificates for the following standards.

• ISO 9001:2008 - certificate

• EN ISO 13485:2003 / AC:2009 - certificate

• EC Directive 98/79/EC - certificate


 

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