BstAP I

Enzyme nameBstAP I
Prototype(ApaBI)
Known analogsApaBI
SKUSE-E259
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'… GCANNNN▼NTGC …3'
3'… CGTN▲NNNNACG …5'
SourceBacillus stearothermophilus AP
Optimal bufferSE-buffer W (10 mM Tris-HCl (pH 8.5 at 25°C); 10 mM MgCl2; 100 mM NaCl; 1 mM DTT.)
Optimal temperature60 °C
Inactivation temperature80 °C
Buffer activity
BGOWYROSE
25257510025100
Unit definitionOne unit of the enzyme is the amount required to hydrolyze 1 μg of Lambda DNA in 1 hour at 60°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; 1 mM DTT; 200 μg/ml BSA; and 50% glycerol Store at -20°C.
LigationAfter 5-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 5 u.a. of enzyme for 16 hours at 60°C.
Methylation sensitivitynot tested
Reagents supplied with enzyme10 X SE-buffer W
NotesHigh enzyme concentration may result in star activity. At 37°C activiti is 50% from maximum. The minimum number of units that resulted in complete digestion of 1 μg of substrate DNA in 16 hours is 0,5.
References
  1. M.A. Abdurashitov, O.A. Belichenko, A.V. Shevchenko, V.S. Dedkov, S.Kh. Degtyarev BstAP I, an ApaB I isoschizomer, cleaves DNA at 5′-GCANNNN^NTGC-3′. // Nucleic Acids Research, Vol. 25, No 12., 2301-2302(1997).
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