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Index / Bioner / dNTP (mixtures) 10mM_1ml(2.5 mMX4) / Product Detail : D-3001 dNTP (mixtures) 10mM_1ml(2.5 mMX4)
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Oct
24th

#D-3001 dNTP (mixtures) 10mM_1ml(2.5 mMX4)

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  Price : 182   EUR
207   USD
141   GBP
768   Zloty
24476   JPY
1410   NOK
1511   SEK
206   CHF

Product name : dNTP (mixtures) 10mM_1ml(2.5 mMX4)

Catalog number : D-3001

Quantity:

Availability: Yes

Supplier name : Bioner

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More Details about

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.



enzyme TOP DNA figure1



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


enzyme TOP DNA figure2
 

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


enzyme TOP DNA figure3
 

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


enzyme TOP DNA figure4



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


enzyme TOP DNA figure5



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

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