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You are here:Home » Molecular Biology » MB-Enzymes, Restriction » Bsu15I (ClaI)

Bsu15I (ClaI)

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Specifications

5'-A T^C G A T-3'
3'-T A G C^T A-5'
Catalog #B3050
Concentration 10u/ul
Source Bacillus subtilis 15
Buffer 33mM Tris-acetate (pH 7.9), 10mM magnesium acetate, 66mM potassium acetate and 0.1mg/ml BSA. Incubate at 37°C
Diluent Buffer 10mM Tris-HCl (pH 7.4 at 25°C), 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA and 50% glycerol. For longer periods–the Storage Buffer should be used.
Storage Buffer 10mM Tris-HCl (pH 8.0 at 25°C), 50mM KCl, 1mM DTT, 0.1mM EDTA, 0.5mg/ml BSA and 50% glycerol.
Overdigestion AssayNo detectable change in the specific fragmentation pattern is observed after 240-fold overdigestion (15u/ug lambda DNA x 16 hours) with Bsu15I.
Ligation/Recutting AssayAfter 50-fold overdigestion (3u/ug DNA x 17 hours) with Bsu15I, more than 95% of the DNA fragments can be ligated at a 5'-termini concentration of 0.15uM. More than 95% of these can be recut.
Labeled Oligonucleotide (LO) AssayNo detectable degradation of a single-stranded and double-stranded labeled oligonucleotide was observed after incubation with 10units of restriction endonuclease for 4 hours.
Blue/White Cloning AssayThe mix of pUC57/HindIII, pUC57/PstI and pUC57/Eco32I digests was incubated with 10units of enzyme for 16 hours. After religation and transformation 0.5% of white colonies were detected.
Methylation EffectsBsu15I cuts neither ATCGm6AT nor ATm5CGAT. Blocked by overlapping Dam methylation. Blocked by CG methylation.
Stability during Prolonged IncubationA minimum of 0.1units of enzyme is required for complete digestion of 1ug of lambda DNA in 16 hours at 37°C.
Thermal InactivationEnzyme is inactivated by incubation at 65°C for 20min.
Digestion of Agarose-embedded DNAA minimum of 5units of enzyme is required for digestion of 1ug of agarose-embedded lambda DNA in 16 hours.
Compatible EndsAciI, Bsp119I, Hin1I, Hin6I, HpaII, MaeII, MspI, NarI, Psp1406I, TaqI, XmiI
Number of Recognition Sites in DNA
Lambda15
PhiX1740
M13mp18/192
pBR3221
pUC18/190
pUC570
pTZ19R/U0
pBluescriptIIKS(-/+)1
pBluescriptIISK(-/+)1
pACYC1771
pACYC1841
Unit DefinitionOne unit is defined as the amount of enzyme required to digest 1ug of lambda DNA dam-in 1 hour at 37°C in 50ul of assay buffer.


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