Recognition site | CTGGAG(N)16↑ GACCTC(N)14↓ |
Source | Bacillus pumilus |
Assayed on | Lambda DNA |
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. |
Optimal SE-buffer | W (10 mM Tris-HCl (pH 8.5 at 25°C); 10 mM MgCl2; 100 mM NaCl; 1 mM DTT.) + BSA |
Enzyme activity (%) | B | G | O | W | Y | Rose | 25 - 50 | 50 - 75 | 75 - 100 | 100 | 50 - 75 | 100 |
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Optimal temperature | 37oC |
Storage conditions | 10 mM Tris-HCl (pH 7.5); 50 mM KCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 200 μg/ml BSA; 50% glycerol. Store at -20°C |
Ligation | After 2-fold overdigestion with enzyme about 95% of the DNA fragments can be ligated and 95% may be recut. |
Non-specific hydrolisis | No nonspecific activity was detected after incubation of 1 μg of Lambda DNA with 2 u.a. of enzyme for 16 hours at 37°C.
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Reagents Supplied with Enzyme |
10 X SE-buffer W, BSA |
Methylation sensitivity | Blocked by overlapping Dcm-methylation (CmCWGG):
5'- C(5mC)TGGAG(N)16-3' 3'- GGA(5mC)CTC(N)14-5' |
Inactivation 20 minutes under | 65oC |
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. |
Quality control | Restriction Endonucleases: Quality Control |
MSDS: | Download MSDS as PDF |
References: | Degtyarev, S.Kh., Morgan P. Unpublished observation (1992).
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