Top DNA Polymerase

Top DNA polymerase는 Thermophile polymerase gene을 DNA recombination 기술을 이용하여 polymerase activity를 향상시켜 cloning한 후, E.coli 에 발현시킨 뒤 고순도로 정제한 94 kDa의 thermostable DNA polymerase로서, PCR법에 의한 DNA의 증폭 및 다양한 증폭실험에 사용할 수 있습니다. Top DNA polymerase의 quality는 activity test, SDS-PAGE / purity test, endonuclease /nickase test를 수행하여 확인하였고, PCR 방법을 사용한 다양한 DNA 증폭 실험에 사용할 수 있습니다.

₩65,000
카탈로그 번호
E-3100-CFG

Features and Benefits

High Yields:
Taq DNA polymerase보다 향상된 증폭 효율을 갖습니다.

High Sensitivity:
Taq DNA polymerase보다 향상된 민감도를 갖습니다.

Stable Reaction:
Top enzyme 에 최적화된 buffer를 제공하여 안정적 반응을 보증합니다.

Reproducibility:
재현성 있는 결과를 위해 바이오니아의 전 제품은 one batch에서 대량생산되고 엄격한 ISO 품질 검사를 통해 항상 균일한 품질을 보증합니다.




Specifications

5' to 3' exonuclease activity: No
3' to 5' exonuclease activity: No
3' – A overhang: Yes
Fragment size: Up to 10 kb

Applications

Real-time quantification of DNA and cDNA targets using dsDNA binding dye
Gene expression profiling
Microbial & viral pathogen detection

Reagents Supplied

10X reaction buffer with (or without) MgCl2: Tris (pH 9.0), 15 mM MgCl2, etc
1X 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

500 U/100 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.

Experimental Data

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 DNA Ladder (D-1030)
Lane 1; 1 ng bacterial genomic DNA
Lane 2; 100 pg bacterial genomic DNA
Lane 3; 10 pg bacterial 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-1030)
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; 3 kb PCR product
Lane 3; 4 kb PCR product
Lane 4; 5 kb PCR product
Lane 5; 6 kb PCR product
Lane 6; 7 kb PCR product
Lane 7; 8 kb PCR product
Lane 8; 9 kb PCR product
Lane 9; 10 kb PCR product

Note : 본 제품에 포함된 Top DNA polymerase는 base incorporation rate를 증가시키기 위하여 point mutation을 통해 5'->3' exonuclease 활성을 제거한 효소입니다. 그러므로 dsDNA binding dye를 이용한 real-time PCR에는 사용하실 수 있지만, probe를 이용하는 real-time PCR에는 사용하실 수 없습니다. Probe를 이용한 qPCR 실험 시에는 AccuPower® Dualstar™ qPCR PreMix를 이용하는 것을 권장합니다.



Cat. No. Product Description Price
D-3001 10 mM dNTP (each 2.5 mM), 1 ml ₩40,000
E-3100 Top DNA Polymerase 500 U, 10 mM dNTPs, 10X reaction buffer with MgCl2 ₩90,000
E-3100-1 Top DNA Polymerase 500 U, 10 mM dNTPs, 10X reaction buffer, 20 mM MgCl2 ₩90,000
E-3100-2 Top DNA Polymerase 500 U, 10X reaction buffer with MgCl2 ₩65,000
E-3100-3 Top DNA Polymerase 500 U, 10X reaction buffer, 20 mM MgCl2 ₩65,000
E-3101 Top DNA Polymerase 2000 U, 10 mM dNTPs, 10X reaction buffer with MgCl2 ₩315,000
E-3101-1 Top DNA Polymerase 2000 U, 10 mM dNTPs, 10X reaction buffer, 20 mM MgCl2 ₩315,000
E-3101-2 Top DNA Polymerase 2000 U, 10X reaction buffer with MgCl2 ₩227,000
E-3101-3 Top DNA Polymerase 2000 U, 10X reaction buffer, 20 mM MgCl2 ₩227,000
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