BglII

Produktübersicht

• Artikelname: BglII
• Artikelnummer: USB-B1087-05
• Hersteller Artikelnummer: B1087-05
• Alternativnummer: USB-B1087-05-2500
• Hersteller: US Biological
• Kategorie: Molekularbiologie

Produktbeschreibung

5-A G A T C T-3 3-T C T A G A-5 Concentration: 10u/ul Source: Bacillus globigii Form, Supplied as aliquid in 50mM Tris-HCl pH 7.5, 10mM MgCl Diluent Buffer: For short term storage (3-4 weeks, dilute with Diluent Buffer (10mM Tris-HCl, pH 7.4,,100mM potassium chloride, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA, 50% glycerol). Storage Buffer: For long term storage, use Storage Buffer (10mM Tris-HCl, pH 8.2, 200mM potassium chloride, 1mM DTT, 0.1mM EDTA, 0.5mg/ml BSA, 50% glycerol). Quality Control: Overdigestion Assay: No detectable change in the specific fragmentation pattern is observed after 160-fold overdigestion (10u/ug lambda DNA x 16 hours) with BglII. Ligation/Recutting Assay: After 50-fold overdigestion (3u/ug DNA x 17 hours) with BglII, more than 95% of the DNA fragments can be ligated at a 5-termini concentration of 0.06uM. 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 10units of BglII for 4 hours. Blue/White Cloning Assay: The mix of pUC57/HindIII, pUC57/PstI and pUC57/Eco32I digests was incubated with 10units of BglII for 16 hours. After religation and transformation <1% of white colonies were detected. Protocol for Digestion: Add: Nuclease free water: 16ul R1625-03: 2ul DNA (0.5-1ug/ml): 1ul B1087-05: 0.5-2ul Mix gently and spin down for a few seconds. Incubate at 37C for 1-16 hours. Protocol for Digestion Directly after Amplification: Add: PCR Reaction Mixture: 10ul (~0.1-0.5ug of DNA) Nuclease free water: 18ul R1625-03: 2ul B1087-05: 1-2ul Mix gently and spin down for a few seconds. Incubate at 37C for 1-16 hours. Enzyme Properties: Methylation Effects: Dam: completely overlaps-no effect. Dcm/CpG/EcoKl: never overlaps-no effect. EcoBl: may overlap-cleavage impaired. Stability during Prolonged Incubation: A minimum of 0.1 units of enzyme is required for complete digestion of 1ug of lambda DNA in 16 hours at 37C. Thermal Inactivation: Enzyme is not inactivated by incubation at 80C for 20 minutes. Digestion of Agarose-embedded DNA: A minimum of 5 units of enzyme is required for digestion of 1ug of agarose-embedded lambda DNA in 16 hours. Compatible Ends: BamHI, BclI, Bsp143I, MboI, PsuI Number of Recognition Sites in DNA: Lambda: 6 PhiX174: 0 M13mp18/19: 1 pBR322: 0 pUC18/19: 0 pUC57: 0 pTZ19R/U: 0