85-1379-51　Eco72I (PmaCI) 2000U　E0374-35

特徴

• Source:E. coli that carries the cloned eco72IR gene from E. coli RFL72
• Concentration:10u/ul
• Unit Definition:One unit is defined as the amount of Eco72l required to digest 1ug of lambda DNA in 1 hour at 37°C in 50ul of reaction buffer.
• 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.
• Diluent Buffer:10mM Tris-HCl, pH 7.4, 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA, 50% glycerol.
• Storage Buffer:10mM Tris-HCl, pH 7.4, 100mM KCl, 1mM DTT, 1mM EDTA, 0.2mg/ml BSA, 50% glycerol.
• Storage and Stability:Store at -20°C. Aliquots are stable for 6 months after receipt.
• Methylation Effects:Dam: never overlaps - no effectDcm: never overlaps - no effectCpG: completely overlaps - blockedEcoKl: may overlap - effect not determinedEcoBl: may overlap - effect not determined
• Number of Recognition Sites in DNA:Lambda: 3, PhiX174: 0, M13mp18/19: 0, pBR322: 0, pUC18/19: 0, pUC57: 0,pTZ19R/U: 0
• Stability during Prolonged Incubation: A minimum of 0.5 units 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 20 minutes
• Star Activity:An excess of Eco72 (15u/ug DNA x 1 hour) or low salt concentration may result in star activity.
• Digestion of Agarose-embedded DNA: A minimum of 5 units of enzyme is required for digestion of 1ug of agarose-embedded lambda DNA in 16 hours.
• Overdigestion Assay:No detectable change in the specific fragmentation pattern is observed after 10-fold overdigestion (10u/ug lambda DNA x 1 hour) with Eco72I (see Star Activity).
• Ligation/Recleavage Assay:The ligation and recleavage assat was replaced with L0 test after validating experiments showed L0 test ability to trace nuclease and phosphatase activities with sensitivity that is highe than L/R by a facrtor of 100.
• Labeled Oligonucleotide (LO) Assay: No detectable degradation of a single-stranded or double-stranded labeled oligonucleotide was observed after incubation with 10 units of Eco72l 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 nuuclease and phosphatase activities with sensitivity that erquals to that of B/W test.