| Recognition site | CAG↑CTG
GTC↓GAC |
| Source | An E.coli strain that carries the cloned Pvu II gene from Proteus vulgaris |
| Assayed on | Lambda DNA |
| Unit definition | One 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-buffer | G (10 mM Tris-HCl (pH 7.6 at 25°C); 10 mM MgCl2; 50 mM NaCl; 1 mM DTT.) + BSA |
| Enzyme activity (%) | | B | G | O | W | Y | Rose | | 25 - 50 | 100 | 25 - 50 | 25 - 50 | 25 - 50 | 100 |
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| Optimal temperature | 37oC |
| Storage conditions | 10 mM Tris-HCl (pH 7.5); 300 mM NaCl; 0,1 mM EDTA; 1 mM DTT; 100 μg/ml BSA; 50% glycerol. Store at -20°C. |
| Ligation | After 10-fold overdigestion with enzyme approximately 70% of the DNA fragments can be ligated and recut. |
| Non-specific hydrolisis | No nonspecific activity was detected after incubation of 1 μg of Lambda DNA with 10 u.a. of enzyme for 16 hours at 37°C.
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| Reagents Supplied with Enzyme |
10 X SE-buffer G, BSA
For E111T and E112T an additional 10X SE-buffer ROSE+ is supplied.
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| Methylation sensitivity | not tested |
| Inactivation 20 minutes under | 80oC |
| Notes | High enzyme concentration may result in star activity.
To obtain 100% activity, BSA should be added to the 1 reaction mix to a final concentration of 100 μg/ml.
Do not use BSA for long incubation.
Turbo Pvu II 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 Pvu II 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 (E111).
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 Pvu II
Incubate at 37°C for 10 min.
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| Quality control | Restriction Endonucleases: Quality Control |
| MSDS: | Download MSDS as PDF |
| References: | Repin, V.E., Degtyarev, S.Kh., Unpublished observations.
 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
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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 |
M - ladder, 1 - Adeno-2 DNA, 2 - Lambda DNA, 3 - T7 DNA, 4 - pUC19, 5 - pBR322
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