Eco72I (PmaCI)

Product Overview

• Article Name: Eco72I (PmaCI)
• Biozol Catalog Number: USB-E0374-35
• Supplier Catalog Number: E0374-35
• Alternative Catalog Number: USB-E0374-35-2000
• Manufacturer: US Biological
• Category: Molekularbiologie

Product Description

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 37C 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 -20C. Aliquots are stable for 6 months after receipt. Methylation Effects: Dam: never overlaps - no effect Dcm: never overlaps - no effect CpG: completely overlaps - blocked EcoKl: may overlap - effect not determined EcoBl: 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 37C. Thermal Inactivation: Enzyme is inactivated by incubation at 65C 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.