Ksp22 I

Enzyme nameKsp22 I
PrototypeBclI
Known analogsBclI, FbaI
SKUSE-E081
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'… T▼GATCA …3'
3'… ACTAG▲T …5'
SourceKurthia species 22
Optimal bufferSE-buffer 2K (10 mM Tris-HCl (pH 7.6 at 25°C); 10 mM MgCl2; 200 mM KCl; 1 mM DTT.)
Optimal temperature37 °C
Inactivation temperature65 °C
Buffer activity
BGOWYROSE
5075505025100
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); 250 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 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 Dam methylation (G m ATC) T GATC A
Reagents supplied with enzyme10 X SE-buffer 2K, 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
// Код для некролога