Product info: Kpn I


Name
Kpn INot blocked by  
GGTAC(5mC) methylation    
Cat. #E079TE080T
Number of reactions100500
Volume, μl100500

Recognition site
GGTACC
CCATGG
SourceAn E.coli strain that carries the cloned Kpn I gene from Klebsiella pneumonia
Assayed onLambda DNA, plasmid DNA
DescriptionTurbo Kpn I is used for short time (15 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.
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); 50 mM NaCl; 0,1 mM EDTA; 1 mM DTT; 200 μg/ml BSA; 50% glycerol; Store at -20°C.
LigationAfter digestion with 1 μl of Turbo Kpn 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 blocked by overlapping Dcm methylation (CmCWGG): GGTACCWGG
Inactivation 20 minutes under
80oC
NotesEnzyme Properties:
1 μl of Turbo Kpn I cuts 1 μg of DNA in 1x SE-Buffer ROSE+ in 15 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 Kpn I
Mix by pipette tip carefully.
Incubate at 37°C for 15 min.
References:Smith, D.I., Blattner, F.R., Davies, J. Nucleic Acids Res. 3: 343-353 (1976).
 Abdurashitov M.A., Tomilov V.N., Chernukhin V.A., Gonchar D. A., Degtyarev S. Kh. Comparative analysis of human chromosomal DNA digestion with restriction endonucleases in vitro and in silico. // "Medical genetics" V.6, No 8, pp 29-36, 2007