BspAC I

Enzyme nameBspAC I
PrototypeAciI
Known analogsAciI, SsiI
SKUSE-E501
Turbo versionNot available
High-concentration versionNot available
Recognition site
5'… C▼CGC …3'
3'… GGC▲G …5'
SourceBacillus species AC
Optimal bufferSE-buffer O (50 mM Tris-HCl (pH 7.6 at 25°C); 10 mM MgCl2; 100 mM NaCl; 1 mM DTT.)
Optimal temperature37 °C
Inactivation temperature65 °C
Buffer activity
BGOWYROSE
10251007510100
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 KH 2 PO 4 (pH 7.2); 100 mM NaCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 200 μg/ml BSA; 50% glycerol. Store at -20°C.
LigationAfter 5-fold overdigestion with enzyme more than 95% of the Lambda DNA fragments can be ligated with T4 DNA Ligase at 16°C and 50% of these can be 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 sensitivityBlocked by CG methylation.
Reagents supplied with enzyme10 X SE-buffer O, BSA
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. BspACI has a non-palindromic recognition site.
References
  1. M.V. Tarasova, Z.K. Dzhanobilova, Y.E. Tomilova, V.S. Dedkov, V.A. Chernukhin, S.Kh. Degtyarev New site-specific endonuclease BspACI from BACILLUS PSYCHRODURANS AC recognizes 5`-C^CGC-3`/3`-GGC^G-5`. // Bulletin of biotechnology and physico-chemical biology named by Yu.A.Ovchinnikov (Moscow), V.5, No.1, p.16-24 (2009). (In Russian)
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