|Source||Bacillus stearothermophilus 31|
|Assayed on||T7 DNA, plasmid DNA|
|Description||Turbo 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.
- Fast DNA analysis
- Fast preparation of vectors for cloning
- Double digestion
|Reaction buffer||SE-buffer ROSE+|
|Storage conditions||10 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.|
|Ligation||After 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 hydrolisis||No 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 sensitivity||Blocked by overlapping Dcm-methylation (CmCWGG): GAGACCWGG|
|Inactivation 20 minutes under||80oC|
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|References:||Shinkarenko, N.M., Lebedeva, N.A., Dedkov, V.S., Abdurashitov, M.A., Degtyarev, S.K. Unpublished observations (1999). |