Eco88I (AvaI)

Produktübersicht

• Artikelname: Eco88I (AvaI)
• Artikelnummer: USB-E0374-45
• Hersteller Artikelnummer: E0374-45
• Alternativnummer: USB-E0374-45-1000
• Hersteller: US Biological
• Kategorie: Molekularbiologie

Produktbeschreibung

5-C Y C G R G 3 3-G R G C Y C 5 Source: E. coli that carries the cloned Eco88IR gene from E. coli RFL88 Unit Definition: One unit is defined as the amount of Eco88I required to digest 1ug of lambda DNA in 1 hour at 37C in 50ul of assay buffer. Storage Buffer: 10mM Tris-HCl, pH 7.4, 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA and 50% glycerol 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 Incubation Temperature: 37C Thermal Inactivation: Eco88I is inactivated by incubation at 65C for 20 minutes Diluent Buffer: 10mM Tris-HCl, pH 7.4, 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA and 50% glycerol Enzyme Properties: Methylation Effects: Dam, Never overlaps - no effect Dcm: Never overlaps - no effect CpG: Completely overlaps - cleavage impaired EcoKI: Never overlaps - no effect EcoBI: Never overlaps - no effect Stability during Prolonged Incubation: A minimum of 0.2units of Eco88I is required for complete digestion of 1ug of lambda DNA in 16 hours at 37C. Digestion of Agarose-embedded DNA: A minimum of 5 units of Eco88I is required for complete digestion of 1ug of agarose-embedded lambda DNA in 16 hours. Compatible Ends: CCCGGG - BshTI, BsaWI, Cfr9I, Cfr10l, Kpn2l, Mrel, NgoMIV, SgAI CTCGAG - Sall, SgrDI, Smol, Xhol Number of Recognition Sites in DNA: Lambda: 8 PhiX174: 1 pBR322: 1 pUC57: 1 pUC18/19: 1 pTZ19R/U: 1 M13mp18/19: 2 Overdigestion Assay: No detectable change in the specific fragmentation pattern is observed after 160-fold overdigestion with Eco88I (10U/ug lambda DNA x 16 hours). Ligation/Recutting Assay: The ligation and recleavage assay was replaced with L0 test after validating experiments showed L0 test ability to trace nuclease and phosphatase activities with sensitivity that is higher than L/R by a factor of 100. Labeled Oligonucleotide (LO) Assay: No detectable degradation of a single-stranded and double-stranded labeled oligonucleotide was observed after incubation with 10 units of Eco88I 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 nuclease and phosphatase activities with sensitivity that equals to that of B/W test. Storage and Stability: May be stored at 4C. For long-term storage, aliquot and store at -20C. Aliquots are stable for 6 months after receipt. For maximum recovery of product, centrifuge the original vial prior to removing the cap. Further dilutions can be made in assay buffer.