Recognition Sequence:


Source: Rhodopseudomonas sphaeroides.

Buffer supplied: 10x Reaction Buffer B2.

Storage buffer: 50 mM KCl, 10 mM Tris-HCl (pH 7.4), 0.1 mM EDTA,
1 mM dithiothreitol, 200 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, 1 mM dithiothreitoland 100 µg/ml BSA.
Incubate at 37°C.

Heat inactivation: 20 minutes at 65°C.

Ligation and recutting: After ten-fold overdigestion with Rsa I,
>95 % of the DNA fragments can be ligated and recut with this enzyme.

DNA Methylation:
No Inhibition: dam, dcm
Inhibition (Blocked by some combinations of overlapping): CpG

Unit Definition: One unit is the amount of enzyme required to completely digest 1 µg of Lambda DNA (113 sites) in 1 hour in a total reaction volume of 50 µl. Enzyme activity was determined in the recommended reaction buffer.