EcoR V

Enzyme nameEcoR V
PrototypeEcoRV
Known analogsEco32I
SKUSE-E059
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'… GAT▼ATC …3'
3'… CTA▲TAG …5'
SourceAn E.coli strain, that carries the cloned gene EcoRV from Escherichia coli
Optimal bufferSE-buffer W (10 mM Tris-HCl (pH 8.5 at 25°C); 10 mM MgCl2; 100 mM NaCl; 1 mM DTT.)
Optimal temperature37 °C
Inactivation temperature80 °C
Buffer activity
BGOWYROSE
025501002550
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 conditions10 mM Tris-HCl (pH 7.5); 50 mM NaCl; 0,1 mM EDTA; 1 mM DTT; 200 μg/ml BSA; 50% glycerol; Store at -20°C.
LigationAfter 20-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 20 u.a. of enzyme fro 16 hours at 37°C.
Methylation sensitivitynot tested
Reagents supplied with enzyme10 X SE-buffer W, BSA
NotesHigh enzyme concentration results in star activity. Do not use BSA for long incubation To obtain 100% activity, BSA should be added to the 1 x reaction mix to a final concentration of 100 μg/ml.
References
  2. External publication Холмина В., Ребентиш Б.A., Скоблов Ю.С., и др. Докл. Акад. Наук СССР 253: 495-497 (1980).
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