AspLE I

Description

Recognition site and hydrolysis position:

Source: Arthrobacter species LE3860
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 O
Enzyme activity (%):

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.
Ligations:
After 10-fold overdigestion with enzyme more than 90% 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 20 u.a. of enzyme for 16 hours at 37°C. Reagents Supplied with Enzyme 10 X SE-buffer O
Reagents Supplied with Enzyme:
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′
Notes:
References:
Vysotskaya, E.M., Gonchar, D.A., Shevchenko, A.V., Abdurashitov, M.A., Degtyarev, S.Kh. Unpublished observations (1998).