BcuI (SpeI)

Produktübersicht

• Artikelname: BcuI (SpeI)
• Artikelnummer: USB-B0808-05
• Hersteller Artikelnummer: B0808-05
• Alternativnummer: USB-B0808-05-400,USB-B0808-05-2000
• Hersteller: US Biological
• Kategorie: Molekularbiologie

Produktbeschreibung

5-A C T A G T-3 3-T G A T C A-5 Concentration: 10u/ul Source: Bacillus coagulans VS 29-022 Unit Definition: One unit is defined as the amount of B0808-05 required to digest 1ug of control DNA fragments in 1 hour at 37C in 50ul of assay buffer. The control DNA is pUC19 DNA with inserted BcuI recognition site - Psp1406I fragments. Storage Buffer: Supplied as a liquid in 10mM Tris-HCl (pH 7.5 at 25C), 300mM KCl, 1mM DTT, 0.1mM EDTA, 0.2mg/ml BSA and 50% glycerol. Supplied with: R1625: Restriction Enzyme Buffer A, 10X: Suplied as a liquid in 33mM Tris-acetate pH 7.9, 10mM magnesium acetate, 66mM potassium acetate, 0.1mg/ml BSA. Incubate at 37C. Diluent Buffer: 10mM Tris-HCl (pH 7.4 at 25C), 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA and 50% glycerol. For longer periods-the Storage Buffer should be used. Overdigestion Assay: No detectable change in the specific fragmentation pattern is observed after 160-fold overdigestion (10u/ug pUC19-BcuI DNA x 16 hours) with BcuI. Ligation/Recutting (L/R) 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 (LO) Assay: No detectable degradation of a single-stranded and double-stranded labeled oligonucleotide was observed after incubation with 10 units of BcuI for 4 hours. Blue/White (B/W) 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. Stability during Prolonged Incubation: A minimum of 0.5units of BcuI is required for complete digestion of 1ug of Ad2 DNA in 16 hours at 37C. Methylation Effects on Digestion: Dam, Dcm, CpG - Never overlaps, no effect EcoKI, EcoBI - effect not determined Digestion of Agarose-embedded DNA: A minimum of 10 units of BcuI is required for complete digestion of 1ug of agarose-embedded Adenovirus-2 DNA in 16 hours. Compatible Ends: Eco130I, NheI, XbaI, XmaJI Number of Recognition Sites in DNA: Ad2: 3 Lambda: 0 PhiX174: 0 M13mp18/19: 0 pBR322: 0 pUC18/19: 0 pUC57: 0 pTZ19R/U: 0 Thermal Inactivation: BcuI is not inactivated by incubation at 80C for 20min. Inactivation Procedure: 1. To prepare the digested DNA for electrophoresis: (a) stop the digestion by adding 0.5M EDTA, pH 8.0, to achieve a 20mM final concentration. Mix thoroughly, add an electrophoresis loading dye and load onto gel. 2. To prepare DNA suitable for further enzymatic reactions: (a) extract with phenol/chloroform, precipitate with ethanol or isopropanol, wash the pellet with 75% cold ethanol and air dry, (b) dissolve DNA in either nuclease-free water, TE buffer, or a buffer suitable for further applications, (c) check the DNA concentration in the solution.