Product info: Pce I


Name
Pce I
Cat. #E105TE106T
Number of reactions50250
Volume, μl50250

Recognition site
AGGCCT
TCCGGA
SourcePlanococcus citreus
Assayed onLambda DNA, plasmid DNA
DescriptionTurbo Pce I is used for short time (10 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
50oC
Storage conditions 10 mM Tris-HCl (pH 7.5); 100 mM NaCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 100 μg/ml BSA; 50% glycerol; Store at -20°C.
LigationAfter digestion with 1 μl of Turbo Pce I, approximately 70% 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 Pce 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 Pce I
Mix by pipette tip carefully.
Incubate at 50°C for 10 min.
Incubation at 37°C results in 50-75% activity.
References:Gonchar, D.A., Shinkarenko, N.M., Dedkov, V.S., Popichenko, D.V., Degtyarev, S.K. Unpublished observations (2001).