Cfr13I (AsuI)

Sequence:\n5'-G^G N C C-3' \n3'-C C N G^G-5'\n\nRestriction Enzyme Buffer A, 10X (R1625): Supplied to simplify buffer selection for double digests.\n\nEnzyme Properties:\nNumber of Recognition Sites in DNA:\nLambda: 74 \nPhiX174: 2\npBR322: 15\npUC57: 8\npUC18/19: 6\npTZ19R/U: 6\nM13mp18/19: 4\n\nMethylation Effects: Cfr13I does not cut GGNCm5C. Blocked by overlapping Dcm and CG methylation.\n\nStability during Prolonged Incubation: A minimum of 0.3units of enzyme is required for complete digestion of 1ug of lambda DNA in 16 hours at 37 degrees C.\n\nThermal Inactivation: Enzyme is inactivated by incubation at 65 degrees C for 20min.\n\nCompatible Ends: CpoI, Eco47I, Psp5II, SanDI: G^G(A/T)CC\n\nQuality Control:\nOverdigestion Assay: No detectable change in the specific fragmentation pattern is observed after 160-fold overdigestion (10u/ug lambda DNA x 16 hours) with Cfr13I. \n\nLigation/Recutting Assay: After 50-fold overdigestion (3u/ug DNA x 17 hours) with Cfr13I, more than 95% of the DNA fragments can be ligated at a 5'-termini concentration of 0.3uM. More than 95% of these can be recut. \n\nLabeled 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.\n\nStorage and Stability:\nMay be stored at 4 degrees C for short-term only. For long-term storage, store at -20 degrees C. Aliquots are stable for at least 6-12months at -20 degrees C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.

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SPECIFICATIONS

Catalog Number

C3300

Size

1000U

Form

Supplied as a liquid in 10mM potassium phosphate, pH 7.5 at 25 degrees C, 100mM potassium chloride, 1mM EDTA, 7mM 2-mercaptoethanol, 0.2mg/ml BSA, 50% glycerol.