MspA1 I

Enzyme nameMspA1 I
PrototypeNspBII
Known analogsNspBII
SKUSE-E191
Turbo versionNot available
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'… CmG▼CKG …3'
3'… GKC▲GmC …5'
SourceMoraxella species A1
Optimal bufferSE-buffer Y (33 mM Tris-acetate (pH 7.9 at 25°C); 10 mM magnesium acetate; 66 mM potassium acetate; 1 mM DTT.)
Optimal temperature37 °C
Inactivation temperature65 °C
Buffer activity
BGOWYROSE
10751025100100
Unit definitionOne 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 onLambda DNA
Storage conditions20 mM Tris-HCl (pH 7.6); 300 mM NaCl; 0,1 mM EDTA; 7 mM 2-mercaptoethanol; 10 mM MgCl2, 200 μg/ml BSA, 50% glycerol. Store at -20°C. Storage at -70°C is recommended for periods longer than 30 days.
LigationAfter 10-fold overdigestion with enzyme 90% of the DNA fragments can be ligated and recut.
Nonspecific hydrolysisNo nonspecific activity was detected after incubation of 1 μg of Lambda DNA with 10 u.a. of enzyme for 16 hours at 37°C.
Methylation sensitivitynot tested
Reagents supplied with enzyme10 X SE-buffer Y, BSA
NotesTo 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
  1. Unpublished data Degtyarev, S.K Unpublished observations.
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