85-1379-35　CpoI (RsrII) 1000u　C7906

特徴

• 5'-C G↓G W C C G-3' 3'-G C C W G↑G C-5'
• Source:Caseobacter polymorphus
• Unit Definition:One unit is defined as the amount of Cpol required to digest 1ug of lambda DNA in 1 hour at 37°C in 50ul of recommended reaction buffer.
• Storage Buffer:10mM Tris-HCl, pH 7.4, 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA and 50% glycerol.
• Supplied with:R1625: Restriction Enzyme Buffer A, 10X:Dilute to 1X for use. 1X buffer composition is 33mM Tris-acetate pH 7.9, 10mM magnesium acetate, 66mM potassium acetate, 0.1mg/ml BSA.
• Incubation Temperature:37ºC
• Thermal Inactivation:Cpol is inactivated by incubation at 65°C for 20 minutes.
• Diluent Buffer:10mM Tris-HCl, pH 7.4, 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA and 50% glycerol.
• Enzyme Properties:Methylation Effects:Dam, Never overlaps - no effectDcm: Never overlaps - no effectCpG: Completely overlaps - blockedEcoKI: Never overlaps - no effectEcoBI: Never overlaps - no effect
• Stability during Prolonged Incubation:A minimum of 0.5units of Cpol is required for complete digestion of 1ug of lambda DNA in 16 hours at 37°C.
• Compatible Ends:Cfr13I, Eco47I, Psp5II, SanDI
• Number of Recognition Sites in DNA:Lambda: 5 PhiX174: 0 M13mp18/19: 0 pBR322: 0 pUC18/19: 0 pUC57: 0 pTZ19R/U: 0
• Overdigestion Assay:No detectable change in the specific fragmentation pattern is observed after 160-fold overdigestion with Cpol (10U/ug lambda DNA x 16 hours).
• Ligation/Recutting Assay:The ligation and recleavage assay was replaced with L0 test after validating experiments showed L0 test ability to trace nuclease and phosphatase activities with sensitivity that is higher than L/R by a factor of 100.
• Labeled Oligonucleotide (LO) Assay:No detectable degradation of a single-stranded and double-stranded labeled oligonucleotide was observed after incubation with 10 units of Cpol for 4 hours.
• Blue/White Cloning Assay:The B/W assay was replaced with L0 test after validating experiments showed L0 test ability to detect nuclease and phosphatase activities with sensitivity that equals to that of B/W test.
• Storage and Stability:May be stored at 4°C. For long-term storage, aliquot and store at -20°C. Aliquots are stable for 6 months after receipt. For maximum recovery of product, centrifuge the original vial prior to removing the cap. Further dilutions can be made in assay buffer.