Eco57I

Product Overview

• Article Name: Eco57I
• Biozol Catalog Number: USB-E0374-30
• Supplier Catalog Number: E0374-30
• Alternative Catalog Number: USB-E0374-30-200,USB-E0374-30-1000
• Manufacturer: US Biological
• Category: Molekularbiologie

Product Description

5-C T G A A G(N)16 -3 3-G A C T T C(N)14 -5 Concentration: 5u/ul Source: E.coli that carries the cloned eco57IR gene from E.coli RFL57 Buffer: [10mM Tris-HCl (pH 7.5), 10mM MgCl2, 50mM NaCl and 0.1mg/ml BSA] + 0.01mM S-adenosylmethionine (supplied). Incubate at 37C Diluent Buffer: 10mM Tris-HCl (pH 7.4 at 25C), 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA and 50% glycerol. For longer periods-the Storage Buffer should be used. Storage Buffer: 10mM potassium phosphate (pH 7.4 at 25C), 100mM NaCl, 1mM EDTA, 7mM 2-mercaptoethanol and 50% glycerol. Overdigestion Assay: No detectable change in the specific fragmentation pattern is observed after 80-fold overdigestion (5u/ug lambda DNA x 16 hours) with Eco57I (see Star Activity). Ligation/Recutting Assay: After 16-fold overdigestion (1u/ug DNA x 16 hours) with Eco57I, approximately 70% of the DNA fragments can be ligated at a 5-termini concentration of 0.2uM. None of these can be recut due to the methylation at the recognition sequence by restriction enzyme. Labeled Oligonucleotide (LO) Assay: No detectable degradation of a single-stranded and double-stranded labeled oligonucleotide was observed after incubation with 10 units of restriction endonuclease for 4 hours. Star Activity: An excess of enzyme (7.5u/ug DNA x 16 hours) may result in star activity. Stability during Prolonged Incubation: A minimum of 1.0 unit of enzyme is required for complete digestion of 1µg of lambda DNA in 16 hours at 37C. Thermal Inactivation: Enzyme is inactivated by incubation at 65C for 20min. Number of Recognition Sites in DNA: Lambda: 40 PhiX174: 0 M13mp18/19: 0 pBR322: 2 pUC18/19: 2 pUC57: 2 pTZ19R/U: 2 pBluescriptIIKS (-/+): 2 pBluescriptIISK (-/+): 2 pACYC177: 1 pACYC184: 2 Unit Definition: One unit is defined as the amount of enzyme required to digest (a maximal cleavage level is achieved at which no change in the fragmentation pattern is observed with further increase of enzyme) 1ug of lambda DNA in 1 hour at 37C in 50ul of assay buffer. Requirements: Only Mg2+ for its activity, but is stimulated by S-adenosylmethionine. 10uM S-adenosylmethionine gives more than a 100-fold increase in Eco57I activity. The cleavage of DNA by Eco57I is never complete.