|Source||An E.coli strain that carries the cloned Pvu II gene from Proteus vulgaris|
|Assayed on||Lambda DNA, plasmid DNA|
|Description||Turbo Pvu II is used for short time (10 min) DNA digestion in universal (ROSE+) SE-Buffer. |
Reaction Original SibEnzyme (ROSE+) Buffer is a modified universal ROSE (Reaction Original SibEnzyme) Buffer, specially designed for Restriction Endonucleases that require BSA to obtain 100% activity. The concentration of BSA in 1X ROSE+ Buffer is 100 μg/ml. ROSE+ Buffer is perfect for DNA cleavage with SE Turbo Restriction Endonucleases and for double digestion. Do not use this Buffer for long incubation (16 hours) with the enzymes having star activity.
- Fast DNA analysis
- Fast preparation of vectors for cloning
- Double digestion
|Reaction buffer||SE-buffer ROSE+|
|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 digestion with 1 μl of Turbo Pvu II, approximately 70% of the DNA fragments can be ligated with high-activity T4 DNA Ligase and recut.|
|Non-specific hydrolisis||No detectable degradation of a single-stranded and double-stranded oligonucleotide was observed after incubation with 1 μl of restriction endonuclease for 3 hours.
|Reagents Supplied with Enzyme||
10 X SE-buffer ROSE+
|Methylation sensitivity||not tested|
|Inactivation 20 minutes under||80oC|
1 μl of Turbo Pvu I I cuts 1 μg of DNA in 1x SE-Buffer ROSE+ in 10 min (assayed on Lambda and plasmid DNA). A short time of DNA digestion requires high quality purification of DNA sample (PCR fragments should be purified after amplification).
Please note that supercoiled plasmid DNA and PCR fragments may have varying rates of cleavage and sometimes need more time to be completely digested.
Standard protocol of Turbo reaction:
20 μl of the reaction volume:
10x SE-Buffer ROSE+ - 2 μl
DNA - 0.2-1 μg
Nuclease-free water - to 20 μl
+ 1 μl of Turbo Pvu II
Mix by pipette tip carefully.
Incubate at 37°C for 10 min.
|References:||Repin, V.E., Degtyarev, S.Kh., Unpublished observations.|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