Description
Recognition site and hydrolysis position:
TTSAA | TTS↑AA |
AASTT | AA↓STT |
Source: Agrococcus species 25
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 Y
Enzyme activity (%):
B | G | O | W | Y | ROSE |
75 | 50 | 10 | 10 | 100 | 50 |
Storage conditions: 10 mM Tris-HCl(pH 7.5); 100 mM KCl; 0,1 mM EDTA; 7 mM 2-mercaptoethanol; 200 μg/ml BSA, 50% glycerol. Store at -20°C.
Ligations:
After 5-fold overdigestion with enzyme 90% of DNA fragments can be ligated and recut.
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 methylation: 5`-TTSA(m6A)-3`/ 3`-(m6A)ASTT-5`.
Notes: To obtain 100% activity, BSA should be added to the 1x reaction mix to a final concentration of 100 μg/ml.
Do not use BSA for long incubation.
References: