Description
Recognition site and hydrolysis position:
TCGCGA | TCG↑CGA |
AGCGCT | AGC↓GCT |
Source: Nocardia rubra
Unit definition:
One unit of the enzyme is the amount required to hydrolyze 1 μg of Lambda DNA (dam-) in 1 hour at 37°C in a total reaction volume of 50 μl.
Assayed on:
Lambda DNA (dam-)
Optimal SE-buffer: SE-buffer W
Enzyme activity (%):
B | G | O | W | Y | ROSE |
0 | 10 | 75 | 100 | 10 | 100 |
Storage conditions: 10 mM Tris-HCl (pH 7.5); 250 mM NaCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 50% glycerol; Store at -20°C.
Ligations:
After 5-fold overdigestion with enzyme about 50% of the DNA fragments can be ligated and 90% of them can be recut. In the presence of 10% PEG ligation is better.
Non-specific hydrolisis:
No nonspecific activity was detected after incubation of 1 μg of Lambda DNA with 10 u.a. of enzyme for 16 hours at 37°C.
Reagents Supplied with Enzyme:
Methylation sensitivity: Blocked by overlapping Dam methylation (GmATC): TCGCGATC.
Notes:
References:
Comb, D.G., Schildkraut, I., Greenough, L. Unpublished observations.