Product info: Hae III


Name
Hae IIINew package    
Cat. #E067E068E068XE067TE068T
Package, u.a.20001000010000200010000
Concentration, u.a./ml1000010000500001000010000

Recognition site
GGCC
CCGG
SourceAn E.coli strain, that carries the cloned Hae III gene from Haemophilus aegyptius
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-bufferG (10 mM Tris-HCl (pH 7.6 at 25°C); 10 mM MgCl2; 50 mM NaCl; 1 mM DTT.)
Enzyme activity (%)
BGOWYRose
75 - 10010025 - 5050 - 7550 - 75100
Optimal temperature
37oC
Storage conditions10 mM Tris-HCl (pH 7.5); 100 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 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 G
For E067T and E068T an additional 10X SE-buffer ROSE is supplied.
Methylation sensitivitynot tested
Inactivation 20 minutes under
80oC
NotesTurbo Hae III 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 Hae III cuts 1 μg of DNA in 1 x SE-Buffer G 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 (E067).
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 Hae III
Incubate at 37°C for 10 min.
Quality control
Restriction Endonucleases: Quality Control
MSDS:Download MSDS as PDF
References:Middleton, J.H., Edgell, M.H., Hutchison, C.A. J. Virol. 10:42-50 (1972).
 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
 M.A. Abdurashitov, V.N. Tomilov, V.A. Chernukhin, D.A. Gonchar, S.Kh. Degtyarev Mammalian chromosomal DNA digestion with restriction endonucleases in silico // "Ovchinnikov bulletin of biotechnology and physical and chemical biology" V.2, No 3, pp 29-38, 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