Hga I

Enzyme name Hga I
Prototype HgaI
Known analogs CseI
SKU SE-E461
Turbo version Not available
High-concentration version Not available
Restriction type Type 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'
Source An E.coli strain that carries the cloned HgaI gene from Haemophilus gallinarum
Optimal buffer SE-buffer B (10 mM Tris-HCl (pH 7.6 at 25°C); 10 mM MgCl2; 1 mM DTT.)
Optimal temperature 37 °C
Inactivation temperature 65 °C
Buffer activity
BGOWYROSE
1007510255050
Unit definition One 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 on DNA pBR322
Storage conditions 10 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.
Ligation After 3-fold overdigestion with enzyme more than 90% of the DNA fragments can be ligated and recut.
Nonspecific hydrolysis Incubation with > 2 units of HgaI per 1 μg of DNA and digestion > 1 hour are not recommended.
Methylation sensitivity Blocked by CG methylation.
Reagents supplied with enzyme 10 X SE-buffer B
Notes Incubation 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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