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Golden Gate Assembly, Restriction Enzyme Digestion

Catalog#: R0580S
LID#: 14113D
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LicensesNew England Biolabs, Inc.
U.S. Patent No. 6,764,843
Reaction Conditions1X NEBuffer 3.1Incubate at 55°C
1X NEBuffer 3.1:100 mM NaCl50 mM Tris-HCl10 mM MgCl2100 µg/ml BSApH 7.9 @ 25°C
Storage Conditions10 mM Tris-HCl300 mM NaCl1 mM DTT0.1 mM EDTA500 µg/ml BSA50% GlycerolpH 7.4 @ 25°C
Heat Inactivation80°C for 20 min
Activity in NEB BuffersNEBuffer 1.1: 10%NEBuffer 2.1: 50%NEBuffer 3.1: 100%CutSmart™ Buffer: 25%
Diluent CompatibilityDiluent B
Methylation Sensitivitydam methylation: Not Sensitivedcm methylation: Not SensitiveCpG Methylation: Blocked
Activity at Temperature@37°C: 20%@55°C: 100%
Quality Control AssaysThe following Quality Control Tests are performed on each new lot and meet the specifications designated for the product. Individual lot data can be found on the Product Summary Sheet/Datacard or Manual which can be found in the Supporting Documents section of this page.
Endonuclease Activity (Nicking):The product is tested in a reaction containing a supercoiled DNA substrate. After incubation for 4 hours the percent converted to the nicked form is determined by agarose gel electrophoresis.Exonuclease Activity (Radioactivity Release):The product is tested in a reaction containing a radiolabeled mixture of single and double-stranded DNA. After incubation for 4 hours the exonuclease activity is determined by the % release of radioactive nucleotides.Ligation and Recutting (Terminal Integrity):After an over-digestion of DNA with a restriction endonuclease the percentage of the DNA fragments ligated with T4 DNA ligase and the percentage that can be recut are determined by agarose gel electrophoresis.Non-Specific DNase Activity (16 hour):The product is tested for non-specific nuclease degradation in a reaction containing a DNA substrate. After incubation for 16 hours there is no detectable degradation of the DNA substrate as determined by agarose gel electrophoresis.
NotesBsmBI is an isoschizomer of Esp3I.May exhibit star activity in NEBuffer 2.1.
Unit DefinitionOne unit is defined as the amount of enzyme required to digest 1 µg of ? DNA in 1 hour at 55°C in a total reaction volume of 50 µl.
SourceAn E.coli strain that carries the BsmBI gene from Bacillus stearothermophilus B61 (Z. Chen).
Unit Size200 units
Concentration10,000 units/ml
Safety StatementsExcept as otherwise agreed in writing by our authorized representative, all products are for RESEARCH USE ONLY, AND NOT FOR HUMAN OR ANIMAL THERAPEUTIC OR DIAGNOSTIC USE.
Sample AvailabilityYes
SynonymRestriction Endonucleases: B, Time-Saver™ Qualified Restriction Enzymes

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Storage Temperature: -20°C

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