Ecl136II (EcoICRI)

Produktübersicht

• Artikelname: Ecl136II (EcoICRI)
• Artikelnummer: USB-E0274
• Hersteller Artikelnummer: E0274
• Alternativnummer: USB-E0274-1500
• Hersteller: US Biological
• Kategorie: Molekularbiologie

Produktbeschreibung

5.....G A G C T C.....3 3.....C T C G A G.....5 Unlike SacI, Ecl136II produces DNA fragments with blunt ends. Source: Enterobacter cloacae RFL136 Concentration: 10 units/ul Unit Definition: One unit is defined as the amount of enzyme required to digest 1ug of lambda DNA-HindIII fragments in 1 hour at 37C in 50ul of assay buffer. 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). Supplied with: R1625-55: Restriction Enzyme Buffer for Ecl136II, 10X: Supplied as a liquid 10mM Bis-Tris Propane-HCl, pH 6.5, 10mM MgCl2, 0.1mg/ml BSA. Incubate at 37C. Diluent and Storage Buffer: 10mM Tris-HCl , pH 7.4 at 25C, 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA, 50% glycerol. Quality Control: Overdigestion Assay: No detectable change in the specific fragmentation pattern is observed after 320-fold overdigestion (20u/ug lambda DNA x 16 hours) with Ecl136II. Ligation/Recutting Assay : After 50-fold overdigestion (3u/ug DNA x 17 hours) with Ecl136II, more than 90% of the DNA fragments can be ligated at a 5-termini concentration of 0.01uM. More than 90% of these can be recut. 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. Blue/White Cloning Assay: pUC57 was digested at a unique site within lacZ gene with 10 units of enzyme for 16 hours. After religation and transformation 1% of white colonies were detected. Methylation Effects: The hemi-methylated sequence GAGCTm5C is cleaved by Ecl136II at a significantly slower rate. Cleavage is blocked when both strands are methylated at positions: 5_ GAGCTm5C_3 3_m5CTCGA G_5 [Overlapping CG methylation may influence DNA cleavage] Stability during Prolonged Incubation: A minimum of 0.2 units of enzyme are required for complete digestion of 1ug 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 2 PhiX174 0 M13mp18/19 1 pBR322 0 pUC18/19 1 pUC57 1 pTZ19R/U 1