AdeI (DraIII)

Produktübersicht

• Artikelname: AdeI (DraIII)
• Artikelnummer: USB-A0859-75
• Hersteller Artikelnummer: A0859-75
• Alternativnummer: USB-A0859-75-500
• Hersteller: US Biological
• Kategorie: Molekularbiologie

Produktbeschreibung

5-C A C N N N G T G-3 3-G T G N N N C A C-5 Source: Alcaligenes denitrificans SS 3-028 Concentration: 10u/ul Incubation Temperature: 37C Diluent Buffer: 10mM Tris-HCl pH 7.4, 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA, 50% glycerol Storage Buffer: Supplied as a liquid in 10mM Tris-HCl pH 7.5, 300mM KCl, 1mM DTT, 0.1mM EDTA, 0.2mg/ml BSA, 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 at 37C, 10mM magnesium acetate, 66mM potassium acetate, 0.1mg/ml BSA. R1625-02: Restriction Enzyme Buffer C, 10X, Dilute to 1X for use. 1X buffer composition is 10mM Tris-HCl pH 7.5 10mM magnesium chloride, 50mM sodium chloride, 0.1mg/ml BSA. Unit Definition: One unit is defined as the amount of AdeI required to digest 1ug of lambda DNA in 1 hour at 37C in 50ul of assay buffer. Enzyme Properties: Star Activity: An excess of AdeI (20u/ug DNA x 1 hour) may result in star activity. Methylation Effects: Dam, Dcm: Never overlaps - No effect. CpG: May overlap - cleavage impaired. EcoKl, EcoBl: May overlap - Effect not determined. Stability during Prolonged Incubation: A minimum of 0.5units of AdeI is required for complete digestion of 1ug of DNA in 16 hours at 37C. Digestion of Agarose-embedded DNA: A minimum of 10units of AdeI is required for complete digestion of 1ug of agarose-embedded lambda DNA in 16 hours. Number of Recognition Sites in DNA: Lambda: 10 PhiX174, pTZ19R/U, M13mp18/19: 1 pBR322, pUC18/19, pUC57: 0 Thermal Inactivation: AdeI is not inactivated by incubation at 80C for 20min (see Protocol for Inactivation Procedure. Quality Control: Overdigestion Assay: No detectable change in the specific fragmentation pattern is observed after 15-fold overdigestion (15u/ug lambda DNA x 1hour) with AdeI (see Star Activity). Ligation and Recleavage (L/R) Assay: After a 10-fold overdigestion (5u/ug DNA x 2 hours) with Adel, more than 80% of the digested DNA fragments can be ligated at a 5-termini concentration of 0.1uM. More than 90% of these sites can be recut. Labeled Oligonucleotide (LO) Assay: No detectable degradation of a single-stranded or double-stranded labeled oligonucleotide occurred during incubation with 10 units of AdeI for 4 hours. Blue/White (B/W) Cloning Assay: A mixture of pUC57/Hindlll, pUC57/Eco32l and pUC57/Pstl digests when incubated with 10 units of Adel for 1 hour. After religation and transformation, the background level of white colonies was <1%. 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.