Product info: Ssp I


Name
Ssp IBlocked by  
A(6mA)TATT methylation.  
Cat. #E041E042E041TE042T
Package, u.a.50025005002500
Concentration, u.a./ml10000100001000010000

Recognition site
AATATT
TTATAA
SourceAn E.coli strain, that carries the cloned gene SspI from Sphaerotilus species
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-bufferK (10 mM Tris-HCl (pH 7.6 at 25°C); 10 mM MgCl2; 100 mM KCl; 1 mM DTT.) + BSA
Enzyme activity (%)
BGOWYRose
75 - 10050 - 7525 - 5050 - 7575 - 100100
Optimal temperature
37oC
Storage conditions10 mM Tris-HCl (pH 7.5); 200 mM NaCl; 0,1 mM EDTA; 1 mM DTT; 200 μg/ml BSA; 50% glycerol. Store at -20°C.
LigationAfter 10-fold overdigestion with enzyme 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 10 u.a. of enzyme for 16 hours at 37°C.
Reagents Supplied with Enzyme 10 X SE-buffer K, BSA
For E041T and E042T an additional 10X SE-buffer ROSE+ is supplied.
Methylation sensitivityBlocked by methylation AmATATT.
Inactivation 20 minutes under
65oC
NotesHigh enzyme concentration may result in star 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 Ssp 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 Ssp I cuts 1 μg of DNA in 1 x SE-Buffer K 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 (E041).
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 Ssp I
Incubate at 37°C for 10 min.
Quality control
Restriction Endonucleases: Quality Control
MSDS:Download MSDS as PDF
References:Schildkraut, I., Grandoni, R. unpublished observations.
 Chernukhin V.A, Abdurashitov M.A., Tomilov V.N., Gonchar D.A., Degtyarev S.Kh. Comparative analysis of mouse chromosomal DNA digestion with restriction endonucleases in vitro and in silico // "Ovchinnikov bulletin of biotechnology and physical and chemical biology" V.3, No 4, pp 19-27, 2007
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