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T4 Polynucleotide Kinase (3` phosphatase-)

No Product Name Cat.# Conc Size Price(₩) Add
1 T4 Polynucleotide Kinase (3` phosphatase-) M009S 10 units/μl 200 units ₩ 110,000 추가
2 T4 Polynucleotide Kinase (3` phosphatase-) M009L 10 units/μl 1,000 units ₩ 440,000 추가
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Product description

T4 polynucleotide kinase (3' phosphatase-) catalyzes the transfer of the terminal phosphate group of ATP to the 5'-hydroxyl terminus of DNA or RNA. It also can catalyze the exchange of 5'-terminal phosphate groups. The enzyme is totally lacking the 3' phosphatase activity.

 

Characteristics

- Molecular weight: 33 kDa

- Reaction temperature: 37

- Heat inactivation: 65 for 10 min

            

Applications

- End-labeling of DNA or RNA

- Phosphorylate the 5'-hydroxyl ends of linkers

- Label 5'-hydroxyl ends of DNA with [γ-32P]-ATP

 

Quality control

- Purity: >99% on SDS-PAGE

- Endonuclease-free

- Exonuclease-free

- Phosphatase-free

            

Unit Definition

One unit is defined as the amount of enzyme catalyzing the incorporation of 1 nmol of acid-insoluble [32P] in 30 minutes at 37.

 

Storage

10 mM Tris-HCl, 50 mM KCl, 1 mM DTT, 0.1 mM EDTA, 50% Glycerol, 0.1 μM ATP, pH 7.4 @ 25, Store at -20

 

10X T4 Polynucleotide Kinase (3’ phosphatase-) Buffer

700 mM Tris-HCl (pH7.6), 100 mM MgCl2, 50 mM DTT

 

Standard reaction conditions

- Radiolabeling at 5’ ends of DNA.

10X T4 Polynucleotide Kinase buffer

2 μl

T4 Polynucleotide Kinase (10 units/μl)

2 μl

[γ-32P]ATP (3,000 Ci/mmol)

6 μl (20 pmol)

Dephosphorylated DNA 5' end (1 to 20 pmol)

1 μl

Distilled water

Up to 20 μl

→ Incubation at 37 for 30 min

→ Terminate reaction by adding 1 μl of 0.5 M EDTA (pH 8.0) or by incubating the reaction mixture at 65 for 20 min.

 

- Phosphorylation of 5’ ends of oligonucleotides

10X T4 Polynucleotide Kinase buffer (+ATP) or 10X T4 DNA Ligase buffer

2 μl

T4 Polynucleotide Kinase (10 units/μl)

1 μl

Oligonucleotide DNA

 Up to 100 pmol

Distilled water

Up to 20 μl

→ Incubation at 37 for 30 min

→ Terminate reaction by adding 1 μl of 0.5 M EDTA (pH 8.0) or by incubating the reaction mixture at 65 for 20 min 

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