Product info: PspX I


Name
PspX I
Cat. #E477TE478T
Number of reactions20100
Volume, μl20100

Recognition site
VCTCGAGB
BGAGCTCV
SourceAn E.coli strain that carries the cloned PspX I gene from Pseudomonas species A1-1
Assayed onLambda DNA (HindIII-digest), plasmid DNA
DescriptionTurbo 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.
Applications:
- Fast DNA analysis
- Fast preparation of vectors for cloning
- Double digestion
Reaction bufferSE-buffer ROSE
Optimal temperature
37oC
Storage conditions10 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.
LigationAfter 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 hydrolisisNo 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 sensitivitynot tested
Inactivation 20 minutes under
80oC
NotesEnzyme Properties:
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)