AspLE I

Enzyme name AspLE I
Prototype HhaI
Known analogs BstHH I, CfoI, HhaI, Hin6I, HinP1I, HspA I
SKU SE-E221
Turbo version Available
High-concentration version Not available
Restriction type Type IIPiType IIP enzymes are classical Type II restriction enzymes with palindromic recognition sequences and symmetric cleavage sites.
Recognition site
5'… GCG▼C …3'
3'… C▲GCG …5'
Source Arthrobacter species LE3860
Optimal buffer SE-buffer O (50 mM Tris-HCl (pH 7.6 at 25°C); 10 mM MgCl2; 100 mM NaCl; 1 mM DTT.)
Optimal temperature 37 °C
Inactivation temperature не инактивируется
Buffer activity
BGOWYROSE
10751005025100
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
Storage conditions 10 mM Tris-HCl (pH 7.5); 100 mM NaCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 100 μg/ml BSA; 50% glycerol. Store at -20°C.
Ligation After 10-fold overdigestion with enzyme more than 90% of the DNA fragments can be ligated and recut.
Nonspecific hydrolysis No nonspecific activity was detected after incubation of 1 μg of Lambda DNA with 20 u.a. of enzyme for 16 hours at 37°C.
Methylation sensitivity Blocked by 5'-G(5mC)GC-3'/3-CG(5mC)G-5' methylation. Not blocked by 5'-GCG(5mC)-3'/3`-(5mC)GCG-5` or 5'-GCG(5mC)-3'/3'-CGCG-5'methylation. Cut hemimethylated site: 5'-G(5mC)GC-3'/3'-CGCG-5'
Reagents supplied with enzyme 10 X SE-buffer O
Notes
References
  1. Unpublished data Vysotskaya, E.M., Gonchar, D.A., Shevchenko, A.V., Abdurashitov, M.A., Degtyarev, S.Kh Unpublished observations (1998).
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