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T4 Polynucleotide Kinase(T4 PNK)

No Product Name Cat.# Conc Size Price(₩) Add
1 T4 Polynucleotide Kinase M005S 10 units/µl 500 units ₩ 60,000 추가
2 T4 Polynucleotide Kinase M005M 10 units/µl 1,000 units ₩ 108,000 추가
3 T4 Polynucleotide Kinase M005L 10 units/µl 2,500 units ₩ 240,000 추가
4 T4 Polynucleotide Kinase M005H 50 units/µl 2,500 units ₩ 240,000 추가
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Product description
T4 Polynucleotide Kinase catalyzes the transfer or exchange of phosphoryl group (Pi) from the r position of ATP to the 5' terminus of single-or double-stranded DNA. Nucleoside 3'- monophosphate can be used as substrate for the transfer or the exchange of phosphoryl groups. T4 Polynucleotide Kinase also catalyzes the removal of phosphate groups from 3'-phosphoryl polynucleotide, deoxynucleoside 3'-monophosphate, and deoxynucleoside 3'-diphosphate.

Characteristics
- Molecular weight: 35 kDa
- Reaction temperature: 37℃
- Heat inactivation: 65℃ for 20 min

Applications
- End labeling of DNA or RNA for radioactive probes and nucleotide sequencing
- Addition of 5'-phosphate for DNA ligation
- Removal of 3'-phosphoryl groups.

Quality control
- Purity: >99% on SDS-PAGE
- Endonuclease-free
- Exonuclease-free
- Phosphatase-free

Unit definition
One unit is defined as the amount of enzyme required to incorporate 1 nmol of r-phosphoryl group of ATP into acidinsoluble materials with 0.26 mM 5'-OH DNA in 30 min at 37℃.

Storage buffer
10 mM Tris-HCl (pH 7.4), 50 mM KCl, 0.1 mM EDTA, 1 mM DTT, 0.1 μM ATP, 50% glycerol

10X T4 Polynucleotide Kinase buffer
700 mM Tris-HCl (pH 7.6), 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 2 pmol)

Distilled water

Up to 20 μl

→ Incubate at 37 for 30min

→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

→ Incubate at 37 for 30min

→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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