Bsa29 I

Enzyme nameBsa29 I
PrototypeClaI
Known analogsBanIII, BseCI, BspDI, BspXI, Bsu15I, BsuTUI, ClaI, ZhoI
SKUSE-E205
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'… AT▼CGAT …3'
3'… TAGC▲TA …5'
SourceBacillus stearothermophilus 29
Optimal bufferSE-buffer G (10 mM Tris-HCl (pH 7.6 at 25°C); 10 mM MgCl2; 50 mM NaCl; 1 mM DTT.)
Optimal temperature37 °C
Inactivation temperature65 °C
Buffer activity
BGOWYROSE
25100505075100
Unit definitionOne unit of the enzyme is the amount required to hydrolyze 1 μg of Lambda DNA (dam-) in 1 hour at 37°C in a total reaction volume of 50 μl.
Assayed onLambda DNA (dam-)
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 overlapping Dam methylation (G m ATC) G ATCGAT C. Blocked by CG methylation.
Reagents supplied with enzyme10 X SE-buffer G, BSA (except E205T and E206T).
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.
References
  1. G.G. Prikhodko, N.A. Petrov, V.E. Repin, S.Kh. Degtyarev Substrate specificity determination of restriction endonuclease Bsu15I. // Izvestia SB SA USSR, Biological sciences series, No.2, 108-109 (1989). (In Russian)
  2. Unpublished data Repin, V.E., Degtyarev, S.K Unpublished observations.
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