EcoRI 10u/ul

Artikelnummer: USB-E0375
Artikelname: EcoRI 10u/ul
Artikelnummer: USB-E0375
Hersteller Artikelnummer: E0375
Alternativnummer: USB-E0375-5X5000
Hersteller: US Biological
Kategorie: Molekularbiologie
5-G A A T T C-3 3-C T T A A G-5 Source: E.coli that carries the cloned ecoRIR gene from Escherichia coli RY13 Supplied With: R1625: Restriction Enzyme Buffer A, 10X: Supplied as a liquid in 33mM Tris-acetate, 10mM magnesium acetate, 66mM potassium acetate, 0.1mg/ml BSA (pH 7.9 at 37C). R1625-03: Restriction Enzyme Buffer D, 10X: Supplied in 50mM Tris-HCl, pH 7.5, 10mM MgCl2, 100mM NaCl, 0.02% Triton X-100, 0.1mg/ml BSA. Concentration: 10u/ul. Unit Definition: One unit is defined as the amount of enzyme required to digest 1ug of lambda DNA in 1 hour at 37C in 50ul of assay buffer. Incubation Temperature: 37C Dilution Buffer: 10mM Tris-HCl, pH 7.4 at 25C, 100mM potassium chloride, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA, 50% glycerol. Storage Buffer: Supplied in 10mM potassium phosphate, pH 7.4, 25C, 300mM NaCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA, 0.15% Triton X-100, 50% glycerol Enzyme Properties: Stability during Prolonged Incubation: A minimum of 0.2u of EcoRI are required for complete digestion of 1ug of lambda DNA in 16 hours at 37C. Thermal Inactivation: EcoRI is inactivated by incubation at 65C for 20min. Digestion of Agarose-Embedded DNA: A minimum of 5u of EcoRI is required for complete digestion of 1ug of agarose-embedded lambda DNA in 16 hours. Compatible Ends: XapI, MunI, TasI Number of Recognition Sites in DNA: Lambda: 5 PhiX174: 0 pBR322, 1 pUC57: 1 pUC18/19: 1 pTZ19R/U: 1 M13mp18/19: 1 Quality Control: Overdigestion Assay: No detectable change in the specific fragmentation pattern is observed after 80-fold overdigestion (5u/ug lambda DNA x 16 hours) with EcoRI. Ligation/Recutting Assay: The ligation and recleavage assay was replaced with LO test after validating experiments showed LO test ability to trace nuclease and phosphatase activities with sensitivity that is higher than L/R by a factor of 100. Labeled Oligonucleotide Assay: No detectable degradation of a single-stranded and double-stranded labeled oligonucleotide was observed after incubation with 10u of EcoRI for 4 hours. Blue/White Cloning Assay: The B/W assay was replaced with LO test after validating experiments showed LO test ability to detect nuclease and phosphatase activities with sensitivity that equals to that of B/W test. Methylation Effects: Dam, Dcm, EcoBl, EcoKl: never overlaps - no effect, CpG: may overlap - cleavage impaired..