Bse8 I

Enzyme nameBse8 I
PrototypeBsaBI
Known analogsBsaBI, BseJI, MamI
SKUSE-E147
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'… GATNN▼NNATC …3'
3'… CTANN▲NNTAG …5'
SourceBacillus species 8
Optimal bufferSE-buffer G (10 mM Tris-HCl (pH 7.6 at 25°C); 10 mM MgCl2; 50 mM NaCl; 1 mM DTT.)
Optimal temperature60 °C
Inactivation temperature80 °C
Buffer activity
BGOWYROSE
25100757550100
Unit definitionOne 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 onLambda DNA
Storage conditions10 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.
LigationAfter 5-fold overdigestion with enzyme about 80% of the DNA fragments can be ligated and recut.
Nonspecific hydrolysisLong incubation is not recommended. Extended incubations and high concentration of the enzyme may result in star activity.
Methylation sensitivityBlocked 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 enzyme10 X SE-buffer G
NotesHigh enzyme concentration (more than 5-fold overdigestion on 1 μg substrate) may result in star activity.
References

No publications found.

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