Product info: Sfr303 I


Name
Sfr303 I
Cat. #E127E128E127TE128T
Package, u.a.1000500010005000
Concentration, u.a./ml10000100001000010000

Recognition site
CCGCGG
GGCGCC
SourceStreptomyces fradiae 303
Assayed onLambda DNA
Unit definitionOne unit of the enzyme is the amount required to hydrolyze 1 μg of Lambda DNA in 1 hour at 37°C in a total reaction volume of 50 μl.
Optimal SE-bufferB (10 mM Tris-HCl (pH 7.6 at 25°C); 10 mM MgCl2; 1 mM DTT.)
Enzyme activity (%)
BGOWYRose
10050 - 7510 - 2510 - 2575 - 100100
Optimal temperature
37oC
Storage conditions10 mM Tris-HCl (pH 7.5); 100 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 90% of the DNA fragments can be ligated and recut.
Non-specific hydrolisisNo nonspecific activity was detected after incubation of 1 μg of Lambda DNA with 20 u.a. of enzyme for 16 hours at 37°C.
Reagents Supplied with Enzyme 10 X SE-buffer B
For E127T and E128T an additional 10X SE-buffer ROSE is supplied.
Methylation sensitivitynot tested
Inactivation 20 minutes under
65oC
NotesTurbo Sfr303 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 Sfr303 I cuts 1 μg of DNA in 1 x SE-Buffer B 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 (E127).
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 Sfr303s I
Incubate at 37°C for 10 min.
Quality control
Restriction Endonucleases: Quality Control
MSDS:Download MSDS as PDF
References:Puchkova, L.I., Selina, A.V., Serov, G.D., Krivopalova, G.N., Rechkunova, N.I., Degtyarev, S.Kh., Unpublished observations.
 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
Theoretical diagrams of DNA digestion by this enzyme for the most known DNA substrates:

To view the fragments length values please point mouse cursor over diagram
Fragment lengths
M12345M

M - ladder, 1 - Adeno-2 DNA, 2 - Lambda DNA, 3 - T7 DNA, 4 - pUC19, 5 - pBR322