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nTaq-Pure (purified from insect cell)

No Product Name Cat.# Conc Size Price(₩) Add
1 nTaq-Pure (purified from insect cell) P025PI 5 units/μl 250 units ₩ 120,000 추가
2 nTaq-Pure (purified from insect cell) P050PI 5 units/μl 500 units ₩ 216,000 추가
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Product description

nTaq-Pure (purified from insect cell) is a highly purified polymerase especially for PCR reactions where freedom from bacterial genomic DNA is essential. A gene encoding the Thermus aquaticus (Taq) DNA polymerase was cloned and expressed in Spodoptera frugiperda (Armyworm), and the enzyme was purified to homogeneity. The purified Taq polymerase (nTaq) has optimal activity at high temperatures (72), which helps amplify secondary-structured regions. nTaq DNA Polymerase has greatly reduced activity of 5'->3' exonuclease activity. As a result, formation of erroneously amplified products caused by the 5'->3' exonuclease activity of wild type polymerase is effectively prevented. In addition, this enzyme has been purified from a completely different organism than bacteria (such as E. coli), so there is no prokaryotic DNA contamination

 

Characteristic

- Absolutely free from prokaryotic DNA contamination.

- Enhanced sensitivity.

- Molecular weight: 94 kDa

- Error rate: 2.4 X 10-5

- Thermal stability: Half life of 40 min at 95

- A-tail formation at 3’ ends of amplified duplex DNA.

 

Applications

- Standard / General PCR

- PCR with bacterial DNA

- E. coli contamination studies

- Microbial (i.e., 16S/23S) screening studies

- Forensic studies

- PCR cloning

- RT-PCR

 

Supplied with

- 10X nTaq-Pure buffer (Mg2+ plus)

- dNTP Mixture (2 mM each)

- Sterile water

 

Quality control

- Purity: >99% on SDS-PAGE

- Endonuclease-free

- Exonuclease-free

- RNase-free

- Inhibitor-free

 

Unit definition

One unit is defined as the amount of enzyme required to incorporate 10 nmol of dNTP into acid insoluble materials in 30 min at 74 in a 50-μl reaction mixture (20 mM Tris-HCl/pH 8.8, 50 mM KCl, 2.5 mM MgCl2, 10 mM β-mercaptoethanol, 12.5 μg of calf thymus DNA).

 

Storage buffer

20 mM Tris-HCl (pH 7.9), 100 mM KCl, 0.1 mM EDTA, 1 mM DTT, 0.5% NP-40, 0.5% Tween-20, 50% glycerol.

 

10X nTaq Buffer (Mg2+ plus)

containing 15 mM MgCl2

 

Standard reaction conditions

- PCR mixturea

10X nTaq Buffer (Mg2+ plus)

2 μl

nTaq-Pure (purified from insect cell) DNA Polymerase(5 unit/μl)

0.2 μl

dNTP Mixture (2 mM each)

2 μl

Template DNAc (0.1~500 ng/μl)

1 μl

Primer 1 (5 pmole/μl)

1 μl

Primer 2 (5 pmole/μl)

1 μl

Sterile water

up to 20 μl

aAssemble the reaction mixture on ice

bAdd the PCR polymerase at the final step

cPlasmid DNA, 0.1 ng-30 ng; genomic DNA, 50 ng-500 ng

 

- PCR cycle

Step

Temperature

Time

Cycle

Initial denaturation

95

2 min

1

Denaturation

95

30 sec

25~35

Annealinga

55~65

30~60 sec

Elongation

72

1 min/kb

Final elongation

72

5 min

1

When cycles are over, keep the reaction at 4; may add 10 mM EDTA until use to prevent DNA degradation

aRecommend annealing temperature is 5 to 10 below the lower Tm of the two primers used

 

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