Description
Recognition site and hydrolysis position:
TTTAAA | TTT↑AAA |
AAATTT | AAA↓TTT |
Source: Deinococcus radiophilus
Unit definition:
One unit of the enzyme is the amount required to hydrolyze 1 μg of Lambda DNA in 1 hour at 37°C in a total reaction volume of 50 μl.
Assayed on:
Lambda DNA
Optimal SE-buffer: SE-buffer G
Enzyme activity (%):
B | G | O | W | Y | ROSE |
75 | 100 | 25 | 75 | 75 | 100 |
Storage conditions: 10 mM Tris-HCl (pH 7.5); 250 mM NaCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 100 μg/ml BSA, 50% glycerol; Store at -20°C.
Ligations:
After 20-fold overdigestion with enzyme about 70% of the DNA fragments can be ligated and recut.
Non-specific hydrolisis:
No nonspecific activity was detected after incubation of 1 μg of Lambda DNA with 40 u.a. of enzyme for 16 hours at 37°C.
Reagents Supplied with Enzyme:
Methylation sensitivity: not tested
Notes: To obtain 100% activity, BSA should be added to the 1 x reaction mix to a final concentration of 100 μg/ml.
Do not use BSA for long incubation.
References:
Purvis, I.J., Moseley, B.E.B., Nucleic Acids Res. 11:5467-5474 (1983).