Product info: BstDE I


Name
BstDE I    
Cat. #E227TE228T
Number of reactions50250
Volume, μl50250

Recognition site
CTNAG
GANTC
SourceBacillus stearothermophilus DE
Assayed onLambda DNA, plasmid DNA
DescriptionTurbo BstDE I is used for a short time (10 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
60oC
Storage conditions10 mM Tris-HCl (pH 7.5); 100 mM NaCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 50% glycerol; Store at -20°C.
LigationAfter digestion with 1 μl of Turbo BstDE 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 BstDE 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 BstDE I
Mix by pipette tip carefully.
Incubate at 60°C for 10 min.
References:Shinkarenko, N.M., Shevchenko, A.V., Dedkov, V.S., Abdurashitov, M.A., Degtyarev, S.K. Unpublished observations (1995).
 Murat A Abdurashitov, Victor N Tomilov, Valery A Chernukhin, S. Kh. Degtyarev A physical map of human Alu repeats cleavage by restriction endonucleases // BMC Genomics 2008, 9:305
 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
 V.A. Chernukhin, M.A. Abdurashitov, V.N. Tomilov, D.A. Gonchar, S.Kh. Degtyarev Comparative restriction enzymes analysis of rat chromosomal DNA in vitro and in silico // "Ovchinnikov bulletin of biotechnology and physical and chemical biology" V.2, No 3, pp 39-46, 2006