Ahl I

Enzyme nameAhl I
PrototypeSpeI
Known analogsBcuI, SpeI
SKUSE-E173
Turbo versionAvailable
High-concentration versionNot available
Restriction typeType IIPiType IIP enzymes are classical Type II restriction enzymes with palindromic recognition sequences and symmetric cleavage sites.
Recognition site
5'… A▼CTAGT …3'
3'… TGATC▲A …5'
SourceAlteromonas haloplanktis SP
Optimal bufferSE-buffer B (10 mM Tris-HCl (pH 7.6 at 25°C); 10 mM MgCl2; 1 mM DTT.)
Optimal temperature37 °C
Inactivation temperatureне инактивируется
Buffer activity
BGOWYROSE
10075252575100
Unit definitionOne unit of the enzyme is the amount required to hydrolyze 1 μg of T7 DNA in 1 hour at 37°C in a total reaction volume of 50 μl.
Assayed onT7 DNA
Storage conditions10 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.
LigationAfter 20-fold overdigestion with enzyme more than 90% of T7 DNA fragments can be ligated and recut.
Nonspecific hydrolysisNo nonspecific activity was detected after incubation of 1 μg of T7 DNA with 40 u.a. of enzyme for 16 hours at 37°C.
Methylation sensitivitynot tested
Reagents supplied with enzyme10 X SE-buffer B, BSA
NotesTo 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
  1. Unpublished data Myakisheva, T.V., Belichenko, O.A., Popichenko, D.V., Dedkov, V.S., Degtyarev, S.Kh Unpublished observations (2000).
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