Description
Recognition Site and Hydrolysis Position:
| GGTCTC(N)1 | GGTCTC(N)1↑ |
| CCAGAG(N)6 | CCAGAG(N)6↓ |
Source: Peribacillus species 31
Definition of Activity Unit:
One unit of activity is defined as the amount of enzyme capable of hydrolyzing 1 µg of T7 DNA in a 50 µL reaction mixture in SE buffer Y at 37°C for 1 hour.
Substrate for Activity Determination:
T7 DNA
Optimal Buffer: SE buffer Y
Enzyme Activity in Various Buffers as a Percentage of Maximum:
| B | G | O | W | Y | ROSE |
| 25-50 | 75-100 | 10-25 | 25-50 | 100 | 75 |
Storage Conditions: 10 mM Tris-HCl (pH 7.5), 200 mM NaCl, 0.1 mM EDTA, 200 µg/mL BSA, 7 mM 2-mercaptoethanol, 50% glycerol; Store at -20°C
Ligase Compatibility:
After hydrolysis with a 10-fold excess of enzyme, more than 90% of DNA fragments are ligated by DNA ligase, and approximately 90% of them can be re-cleaved.
Non-Specific Hydrolysis:
The pattern of specific hydrolysis remains unchanged when treating 1 µg of DNA with 20 units of enzyme for 16 hours at 37°C. BSA is not recommended for prolonged incubation.
Methylation Sensitivity: Blocked 90-95% when overlapping with mCG methylation: 5′-GGTCT(5mC)G-3′ and/or 5′-GAGAC(5mC)G-3′.
Supplied with the Enzyme: 10 x SE Buffer Y, BSA (10 mg/mL).
For Turbo packaging, 10 X SE Buffer ROSE is included.
