Product info: Pst I


Name
Pst I
Cat. #E109TE110T
Number of reactions2001000
Volume, μl2001000

Recognition site
CTGCAG
GACGTC
SourceAn E.coli strain that carries the cloned Pst I gene from Providencia stuartii
Assayed onLambda DNA, plasmid DNA
DescriptionTurbo Pst 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
37oC
Storage conditions10 mM Tris-HCl (pH 7.5); 200 mM NaCl; 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 Pst 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 Pst 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 Pst I
Mix by pipette tip carefully.
Incubate at 37°C for 10 min.
References:Smith, D.I., Blattner, F.R., Davies, J. Nucleic Acids Res. 3: 343-353 (1976).