FauND I

Description

Recognition site and hydrolysis position:

Source: An E.coli strain that carries the cloned FauND I gene from Flavobacterium aquatili ND
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 Y
Enzyme activity (%):

Storage conditions: 10 mM Tris-HCl (pH 7.5); 50 mM KCl; 0,1 mM EDTA; 1mM DTT; 200 μg/ml BSA, 50% glycerol; Store at -20°C.
Ligations:
After 10-fold overdigestion with enzyme 80% of the DNA fragments can be ligated and recut. In the presence of 10% PEG ligation is better.
Non-specific hydrolisis:
No nonspecific activity was detected after incubation of 1 μg of Lambda DNA with 10 u.a. of enzyme for 16 hours at 37°C. Reagents Supplied with Enzyme 10 X SE-buffer Y, BSA
Reagents Supplied with Enzyme:
Methylation sensitivity: not tested
Notes: Sensitive to impurities present in some DNA preparations. For example, DNA purified by standard miniprep procedures is cleaved at lower rates.
To obtain 100% activity, BSA should be added to the 1 x reaction mix to a final concentration of 100 μg/ml.
Do not use BSA for long incubation.
References:
Shevchenko, A.V., Belichenko, O.A., Myakisheva, T.V., Dedkov, V.S., Abdurashitov, M.A., Degtyarev, S.K. Unpublished observations (1995).
Abdurashitov M.A., Tomilov V.N., Chernukhin V.A., Gonchar D. A., Degtyarev S. Kh. Comparative analysis of human chromosomal DNA digestion with restriction endonucleases in vitro and in silico. // Translated from “Medical genetics” V.6, No 8, pp 29-36, 2007