|Source||An E.coli strain that carries the cloned PspX I gene from Pseudomonas species A1-1|
|Assayed on||Lambda DNA (HindIII-digest), plasmid DNA|
|Description||Turbo PspX I is used for short time (15 min) DNA digestion in universal (ROSE) SE-Buffer. |
Reaction Original SibEnzyme (ROSE) Buffer is a specially designed universal reaction buffer for the most Restriction Endonucleases. ROSE Buffer is perfect for DNA cleavage with SE Turbo Restriction Endonucleases and for double digestion.
- 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); 200 mM KCl; 0.1 mM EDTA; 7 mM 2 -mercaptoethanol, 200 μg/ml BSA; 50% glycerol. Store at -20°C.|
|Ligation||After digestion with 1 μl of Turbo PspX I, approximately 90% 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 PspX I cuts 1 μg of DNA in 1x SE-Buffer ROSE in 15 min (assayed on Lambda DNA (HindIII-digest) 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 PspX I
Mix by pipette tip carefully.
Incubate at 37°C for 15 min.
|References:||Gonchar D.A., Abdurashitov M.A., Belichenko O.A., Dedkov V.S., Mezentseva N.V., Tomilova J.E., Degtyarev S.Kh. Bulletin of biotechnology and physico-chemical biology named by Yu.A.Ovchinnikov, No. 1, pp18-23 (2005)|