Csp6I (RsaI)

5'-G^T A C-3' \n3'-C A T^G-5'\nUnlike RsaI, Csp6I produces DNA fragments with a 2-base 5'-extension.\n\nSource:\nCorynebacterium species RFL6\n\nConcentration:\n10u/ul\n\nUnit Definition:\nOne unit is defined as the amount of enzyme required to digest 1ug of lambda DNA in 1 hour at 37 degrees C in 50ul of assay buffer.\n\nStorage Buffer:\nSupplied as a liquid in 10mM Tris-HCl, pH 7.4 at 25 degrees C, 100mM potassium chloride, 1mM DTT, 1mM EDTA, 0.2mg/ml BSA, 50% glycerol.\n\nSupplied With:\nR1625 Restriction Enzyme Buffer A, 10X: Supplied as a liquid in33mM Tris-acetate, 10mM magnesium acetate, 66mM potassium acetate, 0.1mg/ml BSA, pH 7.9 at 37 degrees C. \n\nR1625-01 Restriction Enzyme Buffer B, 10X: Supplied as a liquid in 10mM Tris-HCl, pH 7.5, 10mM magnesium chloride, 0.1mg/ml BSA. Incubate at 37 degrees C\nEnzyme Properties:\nStability during Prolonged Incubation: \nA minimum of 0.1u of Csp6I is required for complete digestion of 1ug of lambda DNA in 16 hours at 37 degrees C.\nThermal Inactivation: Csp6I is inactivated by incubation at 65 degrees C for 20min.\nCompatible Ends: FspBI, NdeI, Tru1I, VspI \nNumber of Recognition Sites in DNA:\nLambda: 113\nPhiX174: 11\npBR322, pUC57, pUC18/19: 3\npTZ19R/U: 2\nM13mp18/19: 19\nMethylation Effects: \nDam, Dcm, EcoKI, EcoBI: Never overlaps-no effect\nCpG: May overlap-no effect\nQuality Control:\nOverdigestion Assay: No detectable change in the specific fragmentation pattern is observed after 320-fold overdigestion (20u/ug lambda DNA x 16 hours) with Csp6I.\nLigation/Recutting Assay: After 50-fold overdigestion (3u/ug DNA x 17 hours) with Csp6I more than 95% of the DNA fragments can be ligated at a 5'-termini concentration of 05uM. More than 95% of these can be recut. \nLabeled Oligonucleotide Assay: No detectable degradation of a single-stranded and double-stranded labeled oligonucleotide was observed after incubation with 10u of Csp6I for 4 hours.

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