Product info: Taq I


Name
Taq INew package    
Cat. #E133E134E133TE134T
Package, u.a.200010000200010000
Concentration, u.a./ml20000200002000020000

Recognition site
TCGA
AGCT
SourceAn E.coli strain, that carries the cloned gene Taq I from Thermus aquaticus
Assayed onLambda DNA
Unit definitionOne unit of the enzyme is the amount required to hydrolyze 1 μg of Lambda DNA in 1 hour at 65°C in a total reaction volume of 50 μl.
Optimal SE-bufferY (33 mM Tris-acetate (pH 7.9 at 25°C); 10 mM magnesium acetate; 66 mM potassium acetate; 1 mM DTT.) + BSA
Enzyme activity (%)
BGOWYRose
50 - 7575 - 10075 - 10050 - 75100100
Optimal temperature
65oC
Storage conditions10 mM Tris-HCl (pH 7.5); 300 mM NaCl; 0,1 mM EDTA; 1 mM DTT; 200 μg/ml BSA; 50% glycerol. Store at -20°C.
LigationAfter 20-fold overdigestion with enzyme more than 95% 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 65°C.
Reagents Supplied with Enzyme 10 X SE-buffer Y, BSA
For E133T and E134T an additional 10X SE-buffer ROSE is supplied.
Methylation sensitivitynot tested
Inactivation 20 minutes under
80oC
NotesTo 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 Taq 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 Taq I cuts 1 μg of DNA in 1 x SE-Buffer Y 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 (E133).
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 Taq I
Incubate at 65°C for 10 min.
Quality control
Restriction Endonucleases: Quality Control
MSDS:Download MSDS as PDF
References:Sato, S., Hutchison, C.A. III, Harris, J.I. Proc. Natl. Acad. Sci. U. S. A. 74: 542-546 (1977).
 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
 V.A. Chernukhin, M.A. Abdurashitov, V.N. Tomilov, D.A. Gonchar, S.Kh. Degtyarev Comparative restriction enzymes analysis of rat chromosomal DNA in vitro and in silico // "Ovchinnikov bulletin of biotechnology and physical and chemical biology" V.2, No 3, pp 39-46, 2006
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