Hga I

Enzyme nameHga I
PrototypeHgaI
Known analogsCseI
SKUSE-E461
Turbo versionNot available
High-concentration versionNot available
Restriction typeType IISiType IIS enzymes recognize asymmetric DNA sequences and cleave outside the recognition site at a defined distance. They are widely used in Golden Gate assembly.
Recognition site
5'… GACGC(N)5▼ …3'
3'… CTGCG(N)10▲ …5'
SourceAn E.coli strain that carries the cloned HgaI gene from Haemophilus gallinarum
Optimal bufferSE-buffer B (10 mM Tris-HCl (pH 7.6 at 25°C); 10 mM MgCl2; 1 mM DTT.)
Optimal temperature37 °C
Inactivation temperature65 °C
Buffer activity
BGOWYROSE
1007510255050
Unit definitionOne unit of the enzyme is the amount required to hydrolyze 1 μg of DNA pBR322 in 1 hour at 37°C in a total reaction volume of 50 μl.
Assayed onDNA pBR322
Storage conditions10 mM Tris-HCl (pH 7.5); 50 mM NaCl; 0.1 mM EDTA; 1 mM DTT; 200 μg/ml BSA; 50% glycerol; Store at -20°C.
LigationAfter 3-fold overdigestion with enzyme more than 90% of the DNA fragments can be ligated and recut.
Nonspecific hydrolysisIncubation with > 2 units of HgaI per 1 μg of DNA and digestion > 1 hour are not recommended.
Methylation sensitivityBlocked by CG methylation.
Reagents supplied with enzyme10 X SE-buffer B
NotesIncubation with more than 2 units of enzyme per 1 μg of DNA for longer than 1 hour at 37°C is not recommended.
References
  1. S.Kh. Degtyarev, V.E. Repin, N.A. Bozhko, S.N. Schelkunov Preparation method of restriction endonuclease Hga I // Soviet Union patent SU1389292 (1985). (In Russian)
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