Bse8 I

Enzyme name Bse8 I
Prototype BsaBI
Known analogs BsaBI, BseJI, MamI
SKU SE-E147
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'… GATNN▼NNATC …3'
3'… CTANN▲NNTAG …5'
Source Bacillus species 8
Optimal buffer SE-buffer G (10 mM Tris-HCl (pH 7.6 at 25°C); 10 mM MgCl2; 50 mM NaCl; 1 mM DTT.)
Optimal temperature 60 °C
Inactivation temperature 80 °C
Buffer activity
BGOWYROSE
25100757550100
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); 100 mM NaCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 100 μg/ml BSA; 50% glycerol. Store at -20°C.
Ligation After 5-fold overdigestion with enzyme about 80% of the DNA fragments can be ligated and recut.
Nonspecific hydrolysis Long incubation is not recommended. Extended incubations and high concentration of the enzyme may result in star activity.
Methylation sensitivity Blocked by overlapping Dam-methylation (GmATC): 5’- GmATC N^NNATC-3’/3’- CTmAG N^NNTAG. Blocked by overlapping (5mC)-methylation: 5’-GATNN^NNAT(5mC)-3’/3’-(5mC)TANN^NNTAG-5’. Cuts hemimethylated sequence: 5’-GATNN^NNAT(5mC)-3’/3’-CTANN^NNTAG-5’.
Reagents supplied with enzyme 10 X SE-buffer G
Notes High enzyme concentration (more than 5-fold overdigestion on 1 μg substrate) may result in star activity.
References

No publications found.
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