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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 |
|
| 2 | T4 Polynucleotide Kinase (3` phosphatase-) | M009L | 10 units/μl | 1,000 units | ￦ 440,000 |
|
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
● Material Safety Data Sheet
- Enzynomics_MSDS_M009_T4 Polynucleotide Kinase (3` phosphatase-)_E
