Ajn I

Description

Recognition site and hydrolysis position:

Source: Acinetobacter johnsonii R2
Unit definition:
One unit of the enzyme is the amount required to hydrolyze 1 μg of Lambda DNA in 1 hour at 55°C in a total reaction volume of 50 μl.
Assayed on:
Lambda DNA
Optimal SE-buffer: SE-buffer Y
Enzyme activity (%):

Storage conditions: 10 mM Tris-HCl (pH 7.5); 100 mM NaCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 200 μg/ml BSA; 50% glycerol; Store at -20°C.
Ligations:
After 3-fold overdigestion with enzyme 95% 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 6 u.a. of enzyme for 16 hours at 55°C. Reagents Supplied with Enzyme 10 X SE-buffer Y
Reagents Supplied with Enzyme:
Methylation sensitivity: Not blocked by dcm methylation: CCWGGNotes: At 37°C activity is about 10% from maximum.
References:
V.S. Dedkov, N.A. Mikhnenkova, L.P. Vlasenko, J.E. Tomilova, J.G.Kashirina, D.A. Gonchar, S.Kh. Degtyarev Restriction Endonuclease AjnI from Acinetobacter johnsonii R2, an Isoschizomer of EcoRII, recognizes 5′- CCWGG-3′ and cleaves dcm-methylated sites // Translated from BIOTEHNOLOGIA (Russia) No. 3, pp. 19-24, 2004