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B0808

BcnI (CauII)

Size

£292.00

SKU:
B0808
Storage Conditions:
-20°C
Supplier:
United States Biological
Extra Details:
G 5'-C C^C G G-3' 3'-G G G^C C-5' C Concentration: 10u/ul Source: Bacillus centrosporus RFL1 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 potassium phosphate (pH 7.4 at 25°C), 200mM NaCl, 1mM EDTA, 7mM 2-mercaptoethanol, 0.2mg/ml BSA and 50% glycerol. Overdigestion Assay: No detectable change in the specific fragmentation pattern is observed after 160-fold overdigestion (10u/ug lambda DNA x 16 hours) with BcnI. Ligation/Recutting Assay: After 10-fold overdigestion (2u/ug DNA x 5 hours) with BcnI, more than 80% of the DNA fragments can be ligated in a reaction mixture containing 20–40u of T4 DNA ligase/1ug of fragments and 10% PEG at a 5'-termini concentration of 0.03uM. More than 95% of these can be recut. Labeled Oligonucleotide (LO) Assay: No detectable degradation of a single-stranded and double-stranded labeled oligonucleotide was observed after incubation with 10units of restriction endonuclease for 4 hours. Methylation Effects: Sequence 5'…Cm5CGGG…3' 3'…GGm5CCC…5' is cleaved by BcnI at a significantly slower rate. CG methylation may influence DNA cleavage. Stability during Prolonged Incubation: A minimum of 0.2units of enzyme is required for complete digestion of 1ug of lambda DNA in 16 hours at 37°C. Thermal Inactivation: Enzyme is inactivated by incubation at 65°C for 20min. Compatible Ends: Bme1390I, SatI. Number of Recognition Sites in DNA: Lambda: 114 PhiX174: 1 M13mp18/19: 4 pBR322: 10 pUC18/19: 7 pUC57: 7 pTZ19R/U: 5 pBluescriptIIKS(-/+): 6 pBluescriptIISK(-/+): 6 pACYC177: 6 pACYC184: 10 Unit Definition: One unit is defined as the amount of enzyme required to digest 1ug of lambda DNA in 1 hour at 37°C in 50ul of assay buffer.
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