Product info: AspLE I


Name
AspLE IBlocked by 
5`-G(5mC)GC-3`/3-CG(5mC)G-5` 
methylation   Not blocked by 
5`-GCG(5mC)-3`/3`-(5mC)GCG-5` or 
5`-GCG(5mC)-3`/3`-CGCG-``
methylation   
Cat. #E221TE222T
Number of reactions50250
Volume, μl50250

Recognition site
GCGC
CGCG
SourceArthrobacter species LE3860
Assayed onLambda DNA, plasmid DNA
DescriptionTurbo AspLE I is used for a short time (15 min) of 1 μg DNA digestion in universal (ROSE) SE-Buffer in 50 μl of reaction mixture.
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); 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 AspLE 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 sensitivityBlocked by
5'-G(5mC)GC-3'/3-CG(5mC)G-5' methylation.
Not blocked by
5'-GCG(5mC)-3'/3`-(5mC)GCG-5` or
5'-GCG(5mC)-3'/3'-CGCG-5' methylation.
Cut hemimethylated site: 5'-G(5mC)GC-3'/3'-CGCG-5'
Inactivation 20 minutes
No
NotesEnzyme Properties:
1 μl of Turbo AspLE 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 AspLE I
Mix by pipette tip carefully.
Incubate at 37°C for 15 min.
References:Vysotskaya, E.M., Gonchar, D.A., Shevchenko, A.V., Abdurashitov, M.A., Degtyarev, S.Kh. Unpublished observations (1998).