单位定义

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).

反应条件

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)

使用浓度

10,000 units/ml

贮存溶液

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

热失活

65°C for 20 minutes