T4 Polynucleotide Kinase (2.500 U)

Name: T4 Polynucleotide Kinase (2.500 U)
Brand: NEB
SKU: M0201L
Price: 232.0 EUR
Availability: InStock

von NEB

232,00 €

SKU M0201L

Anzahl

5' phosphorylation of DNA/RNA for subsequent ligation
End labeling DNA or RNA for probes and DNA sequencing
Removal of 3' phosphoryl groups

Catalyzes the transfer and exchange of P_i from the γ position of ATP to the 5´ -hydroxyl terminus of polynucleotides (double-and single-stranded DNA and RNA) and nucleoside 3´-monophosphates. Polynucleotide Kinase also catalyzes the removal of 3´-phosphoryl groups from 3´-phosphoryl polynucleotides, deoxynucleoside 3´-monophosphates and deoxynucleoside 3´-diphosphates (1).

Product Source

A E. coli strain that carries the cloned T4 Polynucleotide Kinase gene. It is purified by a modification of the method of Richardson (1).

Application Features

End-labeling DNA or RNA for probes and DNA sequencing (2)
Addition of 5´-phosphates to oligonucleotides to allow subsequent ligation
Removal of 3´-phosphoryl groups (3)

Properties & Usage

Unit Definition

One Richardson unit is defined as the amount of enzyme catalyzing the incorporation of 1 nmol of acid-insoluble [³²P] in a total reaction volume of 50 μl in 30 minutes at 37°C in 1X T4 Polynucleotide Kinase Reaction Buffer with 66 µM [γ-³²P] ATP (5 x 10⁶ cpm/µmol) and 0.26 mM 5´-hydroxyl-terminated salmon sperm DNA (1).

Reaction Conditions

1X T4 Polynucleotide Kinase Reaction Buffer
Incubate at 37°C

1X T4 Polynucleotide Kinase Reaction Buffer
70 mM Tris-HCl
10 mM MgCl₂
5 mM DTT
(pH 7.6 @ 25°C)

Usage Concentration

10,000 units/ml

Storage Buffer

10 mM Tris-HCl
50 mM KCl
1 mM DTT
0.1 mM EDTA
50% Glycerol
0.1 µM ATP
pH 7.4 @ 25°C

Heat Inactivation

65°C for 20 min