Eco47I (AvaII)

Catalog Number: USB-E0374-15
Article Name: Eco47I (AvaII)
Biozol Catalog Number: USB-E0374-15
Supplier Catalog Number: E0374-15
Alternative Catalog Number: USB-E0374-15-800
Manufacturer: US Biological
Category: Molekularbiologie
Sequence: 5-G G A C C-3 3-C C T G G-5 Source: Escherichia coli RFL47 Concentration: 10u/ul Storage Buffer: Supplied as a liquid in 10mM Tris-HCl pH 7.4, 100mM KCl, 1mM DTT, 1mM EDTA, 0.2mg/ml BSA, 50% glycerol. Diluent Buffer: 10mM Tris-HCl (pH 7.4 at 25C), 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA, 50% glycerol. Supplied with: R1625: Restriction Enzyme Buffer A, 10X: Supplied as a liquid in 33mM Tris-acetate, pH 7.9 at 37C, 10mM magnesium acetate, 66mM potassium acetate, 0.1mg/ml BSA R1625-04: Restriction Enzyme Buffer E, 10X:, Supplied as a liquid in10mM Tris-HCl, pH 8.5, 10mM MgCl2, 100mM KCl, 0.1mg/ml BSA. Number of Recognition Sites in DNA: Lambda 35 chi174, M13mp18/19 1 pBR322 8 pUC57, pUC18/19, pTZ19R/U 2 Overdigestion Assay: No detectable change in the specific fragmentation pattern is observed after 160-fold overdigestion (10u/ug lambda DNA x 16 hours) with Eco47I. Ligation/Recutting Assay: After 50-fold overdigestion (3u/ug DNA x 17 hours) with Eco47I, more than 95% of the DNA fragments can be ligated at a 5-termini concentration of 0.5uM. More than 95% 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. Methylation Effects: Eco47I never overlaps and has no effect on EcoKl and EcoBl. It never overlaps and has no effect on Dam. Dcm and CpG is overlaped and blocked. Note: Eco47I is blocked by overlapping dcm methylation. To avoid dcm methylation, use a dam, dcm strain such as GM2163. Stability during Prolonged Incubation: A minimum of 0.3 units of enzyme is required for complete digestion of 1ug of lambda DNA in 16 hours at 37C. Compatible Ends: Cfr13I, Cpol, Psp5II, SanDI Thermal Inactivation: Enzyme is inactivated by incubation at 65C for 20min. Incubation Temperature: Incubate at 37C Storage and Stability: May be stored at 4C for short-term only. Aliquot to avoid repeated freezing and thawing.. Store at -20C. Aliquots are stable for 6 months after receipt at -20C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.