Product info: Bso31 I


Name
Bso31 Inew package  Blocked by overlapping Dcm-methylation  Not blocked by
  GGTCT(5mC) methylation  
Cat. #E285TE286T
Number of reactions40200
Volume, μl40200

Recognition site
GGTCTC(N)1
CCAGAG(N)5
SourceBacillus stearothermophilus 31
Assayed onT7 DNA, plasmid DNA
DescriptionTurbo Bso31 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 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
55oC
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 Bso31 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 overlapping Dcm-methylation (CmCWGG): GAGACCWGG
Inactivation 20 minutes under
80oC
NotesEnzyme Properties:
1 μl of Turbo Bso31 I cuts 1 μg of DNA in 1x SE-Buffer ROSE in 10 min (assayed on T7 DNA 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 Bso31 I
Mix by pipette tip carefully.
Incubate at 55°C for 10 min.
References:Shinkarenko, N.M., Lebedeva, N.A., Dedkov, V.S., Abdurashitov, M.A., Degtyarev, S.K. Unpublished observations (1999).