Apa I

Enzyme nameApa I
PrototypeApaI
Known analogsBsp120I, PspOM I
SKUSE-E019
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'… GGGCC▼C …3'
3'… C▲CCGGG …5'
SourceAn E.coli strain that carries the cloned Apa I gene from Acetobacter pasteurianus
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
50250010050
Unit definitionOne unit of the enzyme is the amount required to hydrolyze 1 μg of Lambda DNA (dam-dcm-, BamHI-digest) in 1 hour at 37°C in a total reaction volume of 50 μl.
Assayed onLambda DNA/BamHI
Storage conditions10 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.
LigationAfter 20-fold overdigestion with enzyme > 95% of the DNA fragments can be ligated and recut.
Nonspecific hydrolysisNo nonspecific activity was detected after incubation of 1 μg of Lambda DNA with 40 u.a. of enzyme for 16 hours at 37°C.
Methylation sensitivityBlocked by overlapping Dcm methylation(C m CWGG) GGGC CC WGG
Reagents supplied with enzyme10 X SE-buffer Y, BSA (except E019T and E020T).
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. External publication Seurinck, J., Van de Voorde, A., Van Montagu, M. Nucleic Acids. Res.11: 4409-4415 (1983).
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