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Polymerases

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nTaq

No Product Name Cat.# Conc Size Price(₩) Add
1 nTaq (Mg2+ plus buffer) P025A 5 unit/μl 250 units ₩ 60,000 추가
2 nTaq (Mg2+ free buffer) P025B 5 unit/μl 250 units ₩ 60,000 추가
3 nTaq (Mg2+ plus buffer) P050A 5 unit/μl 500 units ₩ 108,000 추가
4 nTaq (Mg2+ free buffer) P050B 5 unit/μl 500 units ₩ 108,000 추가
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Product description

A gene encoding the Thermus aquaticus (Taq) DNA polymerase was cloned and expressed in E. coli, 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, high GC-content or secondary-structured regions can be efficiently amplified due to the genetic modifications in the catalytic domain of the polymerase.

 

Characteristics

- 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

- Amplification of DNA fragments shorter than 3 kb (Suitable for general PCR analysis)

- Amplification of cDNA and genomic DNA.

- Primer extension

- Colony PCR

- Labeling of DNA fragments with radioactive-isotopes

- Nucleotide sequencing

 

Quality control

- Purity: >99% on SDS-PAGE

- Endonuclease-free

- Exonuclease-free

- RNase-free

- Inhibitor-free

 

Supplied with

nTaq (Mg2+ plus buffer)

- 10X nTaq buffer (Mg2+ plus)

- dNTP Mixture (2 mM each)

- Sterile water

nTaq (Mg2+ free buffer)

- 10X nTaq buffer (Mg2+ free)

- dNTP Mixture (2 mM each)

- 25 mM MgCl2

- Sterile water

 

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 buffer: containing 15 mM MgCl2

Mg2+ free buffer: MgCl2 free 

 

Standard PCR conditions

- PCR mixturea

10X nTaq Buffer (Mg2+ plus)

2 μl

nTaq DNA Polymeraseb (5 units/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 mixture at 4; may add 10 mM EDTA until use to prevent DNA degradation.

aRecommended annealing temperatures is 5 to 10 below the lower Tm of the two primers used.

 

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