BstAP I

Enzyme name BstAP I
Prototype (ApaBI)
Known analogs ApaBI
SKU SE-E259
Turbo version Not available
High-concentration version Not available
Restriction type Type 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'
Source Bacillus stearothermophilus AP
Optimal buffer SE-buffer W (10 mM Tris-HCl (pH 8.5 at 25°C); 10 mM MgCl2; 100 mM NaCl; 1 mM DTT.)
Optimal temperature 60 °C
Inactivation temperature 80 °C
Buffer activity
BGOWYROSE
25257510025100
Unit definition One 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 on Lambda DNA
Storage conditions 10 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.
Ligation After 5-fold overdigestion with enzyme more than 90% of the DNA fragments can be ligated and recut.
Nonspecific hydrolysis No 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 sensitivity not tested
Reagents supplied with enzyme 10 X SE-buffer W
Notes High 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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