AspS9 I

Enzyme nameAspS9 I
PrototypeSau96I
Known analogsAsuI, Cfr13I, PspPI, Sau96I
SKUSE-E117
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'… G▼GNCC …3'
3'… CCNG▲G …5'
SourceArthrobacter species S9
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 temperature65 °C
Buffer activity
BGOWYROSE
5050751005075
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 KCl; 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 30 u.a. of enzyme for 16 hours at 37°C.
Methylation sensitivityBloked by overlapping Dcm methylation(C m CWGG) GGN CC WGG
Reagents supplied with enzyme10 X SE-buffer W
References
  1. V.S. Dedkov Analysis of Specificity of DNA-Methyltransferase M.AspS9I in Cell Lysate by means of Restriction Endonucleases Blocking. // Biotekhnologia (Moscow), No 4, p.30-39 (2009). (In Russian)
  4. Unpublished data Dedkov, V.S., Kileva, E.V., Shevchenko, A.V., Degtyarev, S.Kh Unpublished observations (1995).
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