AluB I

Description

Recognition site and hydrolysis position:

Source: Arthrobacter luteus B
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 B
Enzyme activity (%):

Storage conditions: 10 mM Tris-HCl (pH 7.5); 200 mM NaCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 200 μg/ml BSA; 50% glycerol; Store at -20°C.
Ligations:
After 10-fold overdigestion with enzyme 80% of the Lambda DNA fragments can be ligated with T4 DNA Ligase and can be 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 B, BSA
Reagents Supplied with Enzyme:
Methylation sensitivity: AluB I is able to cleave some methylated DNA substrates.
Notes: AluB I is an isoschizomer of Alu I.
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.
References:
V.A. Chernukhin, A. A. Boltengagen, G. V. Tarasova, V.S. Dedkov, S.Kh. Degtyarev New Restriction Endonuclease AluBI from Arthrobacter luteus B – AluI Isoshizomer, nonsensitive to Presence of 5-methylcytosine in the Recognition Sequence AGCT // Translated from “Ovchinnikov bulletin of biotechnology and physical and chemical biology” V.3, No 1, pp 21-27, 2007