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nTaq-HOT (UDG plus)

No Product Name Cat.# Conc Size Price(₩) Add
1 nTaq-HOT (UDG plus) P725U 5 units/μl 250 units ₩ 82,000 추가
2 nTaq-HOT (UDG plus) P750U 5 units/μl 500 units ₩ 150,000 추가
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Product description

nTaq-HOT (UDG plus) is a hot start PCR polymerase which remains inactive at temperatures lower than 75. Nonspecific elongation of incorrectly annealed primers before initiation of PCR reaction is one major cause of nonspecific DNA amplification. For this reason, amplification of nonspecific bands can be effectively prevented by using nTaq-HOT, thus increasing the specificity of target DNA amplification. Also, UDG and dUTP are included in the mixture to prevent the reamplification of carry-over PCR products between reactions.

 

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.

- No activity at temperatures lower than 75. Heating up to 95 results in activation of enzyme.

 

Applications

- High specific amplification of DNA fragments shorter than 3kb.

- Amplification of cDNA and genomic DNA.

- Amplification of template DNA with secondary or higher ordered structure that is resistant to PCR amplification

- Well-suited for an automated PCR machines, for which PCR reaction mixtures are prepared at room

  temperature

- Primer extension

 

Supplied with

- 10X nTaq-HOT buffer

- dNTP Mixture with dUTP (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, 1mM DTT, 0.5% NP-40, 0.5% Tween-20, 50% glycerol.

 

10X nTaq-HOT buffer

Containing 15 mM MgCl2

 

Cautions

nTaq-HOT (UDG plus) requires 10 minutes of initial denaturation to ensure effective activation of the enzyme.

 

Standard reaction conditions

- PCR mixturea

10X nTaq-HOT buffer

μl

nTaq-HOT (UDG plus) DNA polymeraseb (5 units/μl)

0.2 μl

dNTP mixture with UTP (2 mM each)

μl

Template DNA(0.1~500 ng/μl)

μl

Primer 1 (5 pmole/μl)

μl

Primer 2 (5 pmole/μl)

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

Pre-incubation (for UDG)

25

10 min

1

Initial denaturation

95

10 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.

 

● Material Safety Data Sheet

- Enzynomics_MSDS_P725U_nTaq-HOT (UDG plus)

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