Product info: Sfi I


Name
Sfi IBlocked by 
 GGC(5mC)NNNNNGGCC methylation  Not blocked by 
GGCCNNNNNGGC(5mC) methylation  
Cat. #E123E124E123XE124XE123TE124T
Package, u.a.100050001000500010005000
Concentration, u.a./ml100001000040000400001000010000

Recognition site
GGCCNNNNNGGCC
CCGGNNNNNCCGG
SourceAn E.coli strain, that carries the cloned gene Sfi I from Streptomyces fimbriatus
Assayed onT7 DNA
Unit definitionOne unit of the enzyme is the amount required to hydrolyze 1 μg of T7 DNA in 1 hour at 50°C in a total reaction volume of 50 μl.
Optimal SE-bufferG (10 mM Tris-HCl (pH 7.6 at 25°C); 10 mM MgCl2; 50 mM NaCl; 1 mM DTT.) + BSA
Enzyme activity (%)
BGOWYRose
75 - 10010025 - 5025 - 5025 - 5075
Optimal temperature
50oC
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 -20°C.
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 hydrolisisNo nonspecific activity was detected after incubation of 1 μg of T7 DNA with 20 u.a. of enzyme for 16 hours at 50°C.
Reagents Supplied with Enzyme 10 X SE-buffer G, BSA
For E123T and E124T an additional 10X SE-buffer ROSE+ is supplied.
Methylation sensitivityBlocked by overlapping Dcm methylation (CmCWGG): GGCCWGGNNGGCC.
Not blocked by overlapping Dcm methylation (CmCWGG): GGCCNNNNNGGCCWGG.
Inactivation 20 minutes under
65oC
NotesIncubation at 37°C rusults in 75-100% activity.
To obtain 100% activity, BSA should be added to the 1 x reaction mix to a final concentration of 100 μg/ml.
Do not use BSA for long incubation.
Turbo Sfi I can be used for short time (10 min) DNA digestion as well as for standard reaction. The reaction can be performed using optimal or universal (ROSE+) Buffer
Turbo DNA Digestion:
Applications:
-Fast DNA analysis
-Fast preparation of vectors for cloning
-Double digestion
Enzyme Properties: 1 μl of Turbo Sfi I cuts 1 μg of DNA in 1 x SE-Buffer G with BSA or universal 1 x SE-Buffer ROSE+ in 10 min (see the protocol below). A short time of DNA digestion requires high quality purification of DNA sample. The enzyme may be used in standard conditions of DNA cleavage (E123).
Turbo reaction protocol: 20 μl of the reaction volume:
10 x SE Buffer ROSE+ - 2 μl
DNA - 0,2-1 μg
Nuclease-free water - to 20 μl
+ 1μl of Turbo Sfi I
Incubate at 37°C for 10 min.
Quality control
Restriction Endonucleases: Quality Control
MSDS:Download MSDS as PDF
References:Qiang, B.-Q., Schildkraut, I. Nucleic Acids Res. 12: 4507-4515 (1984).
 Dedkov V.S., Degtyarev S. Kh. The resctriction endonucleases detection in colonies of microorganisms Streptomyces and Nocardia // APPLIED BIOCHEMISTRY and MICROBIOLOGY (Russia) Vol 28, 1992, #. 2, 309- 313