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SibEnzyme Ltd. :: Display product information: Sfi I
Name
Sfi I
Catalog numberE123E124E123XE124X
Package, u.a.1000500010005000
Concentration, u.a./ml10000100004000040000

Recognition siteGGCCNNNN^NGGCC
CCGGN^NNNNCCGG
Methylation sensitivityBlocked by overlapping Dcm methylation (CmCWGG): GGCCWGGNNGGCC.
Not blocked by overlapping Dcm methylation (CmCWGG): GGCCNNNNNGGCCWGG.
OriginE.coli strain, that carries the cloned gene Sfi I from Streptomyces fimbriatus
Storage conditions10 mM Tris-HCl (pH 7.5); 200 mM NaCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 200 μg/ml BSA; 50% glycerol. Store at -20C.
Assayed onT7 DNA
LigationAfter 10-fold overdigestion with enzyme more than 70% of the DNA fragments can be ligated and recut. In the presence of 10% PEG ligation is better.
Non-specific activityNo nonspecific activity was detected after incubation of 1 ug of T7 DNA with 20 u.a. of enzyme for 16 hours at 50C.
Optimum temperature50 oC
Inactivation 20 minutes65oC
Optimum SE-bufferG (10 mM Tris-HCl (pH 7.6 at 25C); 10 mM MgCl2; 50 mM NaCl; 1 mM DTT.) + BSA

Enzyme % activity:
BGOWY
75 - 10010025 - 5025 - 5025 - 50

Note: To obtain 100% activity, BSA should be added to the 1 x reaction mix to a final concentration of 100 ug/ml.
Do not use BSA for long incubation.
References: Qiang, B.-Q., Schildkraut, I. Nucleic Acids Res. 12: 4507-4515 (1984).