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Specifications

G
5'-C C^C G G-3'
3'-G G G^C C-5'
C
Catalog #	B0808
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.