Cfr10I (BsrFI)

Sequence: 5'- R ^C C G G Y -3' 3'- Y G G C C^ R -5' Source: E.coli that carries the cloned cfr10IR gene from Citrobacter freundii RFL10 Concentration: 10 units/ul Unit Definition: One unit is defined as the amount of Cfr10I required to digest 1ug of lambda DNA in 1 hour at 37 degrees C in 50ul of assay buffer. Incubation Temperature: 37 degrees C Diluent Buffer: 10mM Tris-HCl (pH 7.4 at 25 degrees C), 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA and 50% glycerol. Storage Buffer: Supplied as a liquid in 10mM potassium phosphate (pH 7.4 at 25 degrees C), 100mM KCl, 1mM DTT, 1mM EDTA, 0.2mg/ml BSA and 50% glycerol. Supplied with: R1625-45: Restriction Enzyme Buffer for Cfr10I, 10X: Supplied as a liquid in 100mM Tris-HCl, pH7.5, 50mM magnesium chloride, 1M sodium chloride, 10mg/ml BSA, 0.2% Triton X-100. R1625: Restriction Enzyme Buffer A, 10X: Supplied as a liquid in 330mM Tris-acetate, pH 7.9, 100mM magnesium acetate, 660mM potassium acetate, 1mg/ml BSA. Enzyme Properties: Star Activity: An excess of Cfr10I (10u/ug DNA x 1 hour) or low salt concentration may result in star activity. Compatible Ends: BshTI, BsaWI, Cfr9I, Eco88I, Kpn2I, MreI, NgoMIV, SgrAI. Methylation Effects: Dam, Dcm: Never overlaps - No effect. CpG: Completely overlaps - Blocked. EcoKI, EcoBI: May overlap - No effect. Stability during Prolonged Incubation: A minimum of 0.1units of Cfr10I is required for complete digestion of 1ug of lambda DNA in 16 hours at 37 degrees C. Digestion of Agarose-embedded DNA: A minimum of 5units of Cfr10I is required for digestion of 1ug of agarose-embedded lambda DNA in 16 hours. Number of Recognition Sites in DNA: Lambda: 61 PhiX174: 0 M13mp18/19, pUC18/19, pUC57: 1 pTZ19R/U: 2 pBR322: 7 Note: For cleavage with Cfr10I, at least two copies of its recognition sequence are required. Thermal Inactivation: Cfr10I is not inactivated by incubation at 80 degrees C for 20min (see Protocol for Inactivation Procedure).