BstHH I

Enzyme nameBstHH I
PrototypeHhaI
Known analogsAspLE I, CfoI, HhaI, Hin6I, HinP1I, HspA I
SKUSE-E143
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'… GCG▼C …3'
3'… C▲GCG …5'
SourceBacillus stearothermophilus HH
Optimal bufferSE-buffer Y (33 mM Tris-acetate (pH 7.9 at 25°C); 10 mM magnesium acetate; 66 mM potassium acetate; 1 mM DTT.)
Optimal temperature50 °C
Inactivation temperatureне инактивируется
Buffer activity
BGOWYROSE
75502550100100
Unit definitionOne unit of the enzyme is the amount required to hydrolyze 1 μg of Lambda DNA in 1 hour at 50°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 50-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 DNA with 100 u.a. of enzyme for 16 hours at 65°C.
Methylation sensitivityBlocked by 5`-G(5mC)GC-3`/3-CG(5mC)G-5` or 5`-G(5mC)GC-3`/3`-CGCG-5`methylation. Not blocked by 5`-GCG(5mC)-3`/3`-(5mC)GCG-5` or 5`-GCG(5mC)-3`/3`-CGCG-5`methylation.
Reagents supplied with enzyme10 X SE-buffer Y, BSA
NotesTo obtain 100% activity, BSA should be added to the 1 x reaction mix to a final concentration of 100 μg/ml Do not use BSA for long incubation.
References
  1. Unpublished data Shapovalova, M.A., Abdurashitov, M.A., Tumanova, I.Y., Dedkov, V.S., Popichenko, D.V., Degtyarev, S.K Unpublished observations (2001)
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