Rga I

Enzyme nameRga I
PrototypeSgfI
Known analogsAsiS I, SgfI
SKUSE-E491
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'… GCGAT▼CGC …3'
3'… CGC▲TAGCG …5'
SourceRhizoblum galegae
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 temperature55 °C
Inactivation temperature80 °C
Buffer activity
BGOWYROSE
75501025100100
Unit definitionOne unit of the enzyme is the amount required to hydrolyze 1 μg of Adenovirus-2 DNA in 1 hour at 55°C in a total reaction volume of 50 μl.
Assayed onAdenovirus-2 DNA
Storage conditions10 mM Tris-HCl (pH 7.5); 250 mM NaCl; 0,1 mM EDTA; 7 mM 2-mercaptoethanol; 50% glycerol. Store at -20°C.
LigationAfter 5-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 Ad2 DNA with 5 u.a. of enzyme for 16 hours at 55°C.
Methylation sensitivityNot blocked by overlapping Dam-methylation (G m ATC) GC GATC GC Blocked by CpG methylation.
Reagents supplied with enzyme10 X SE-buffer Y
NotesHigh enzyme concentration may result in star activity.
References

No publications found.

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