Recognition Sequence:


Source: Streptomyces phaeochromogenes.

Buffer supplied: 10x Reaction Buffer B2.

Storage buffer: 100 mM KCl, 10 mM Tris-HCl (pH 7.4), 0.1 mM EDTA,
1 mM dithiothreitol, 400 µg/ml BSA and 50 % [v/v] glycerol.
Store at -20°C.

Reaction conditions: 50 mM NaCl, 10 mM Tris-HCl (pH 7.9 at 25°C), 10 mM MgCl₂, 1mM dithiothreitol and 100 µg/ml BSA.
Incubate at 37°C.

Heat inactivation: 20 minutes at 65°C.

Ligation and recutting: After ten-fold overdigestion with Sph I,
>98 % of the DNA fragments can be ligated and recut with this enzyme.

DNA Methylation: No Inhibition: dam, dcm, CpG

Unit Definition: One unit is the amount of enzyme required to completely digest 1 µg of Lambda DNA (1 site) in 1 hour in a total reaction volume of 50 µl. Enzyme activity was determined in the recommended reaction buffer.