BfuI (BciVI)

Produktübersicht

• Artikelname: BfuI (BciVI)
• Artikelnummer: USB-B1086-05
• Hersteller Artikelnummer: B1086-05
• Alternativnummer: USB-B1086-05-100
• Hersteller: US Biological
• Kategorie: Molekularbiologie

Produktbeschreibung

Sequence: 5-G T A T C C (N)6 -3 3-C A T A G G (N)5 -5 Source: Bacillus firmus Auk Concentration: 5u/ul Overdigestion Assay: No detectable change in the specific fragmentation pattern is observed after 5-fold overdigestion (5u/ug lambda DNA x 1 hour) with BfuI. Ligation/Recutting Assay: After 5-fold overdigestion (5u/ug DNA x 1 hour) with BfuI, more than 80% of the DNA fragments can be ligated in a reaction mixture containing 20-40u of T4 DNA ligase/1ug of fragments and 10% PEG at a 5-termini concentration of 0.25uM. More than 90% 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 BfuI for 4 hours. Star Activity: A large excess of BfuI (7.5u/ug DNA x 1 hour) may result in star activity. Stability during Prolonged Incubation: A minimum of 1.0unit of BfuI is required for complete digestion of 1ug of lambda DNA in 16 hours at 37C. Digestion of Agarose-embedded DNA: A minimum of 5units of BfuI is required for complete digestion of 1ug of agarose-embedded lambda DNA in 16 hours. Recommended Protocol for Digestion: Add: Nuclease free water: 16ul R1625-20: 2ul DNA (0.5-1ug/ul): 1ul Bful: 0.5-2ul Mix gently, spin down for a few seconds. Incubate at 37C for 1-16 hours. Protocol for Digestion of PCR products directly after amplification: Add: PCR Reaction Mixture: 10ul (~0.1-0.5ug of DNA) Nuclease free water: 18ul R1625-20: 2ul Bful: 1-2ul Mix gently, spin down for a few seconds. Incubate at 37C for 1-16 hours. Unit Definition: One unit is defined as the amount of BfuI required to digest 1ug of lambda DNA dcm in 1 hour at 37C in 50ul of reaction buffer. Supplied With: R1625: Restriction Enzyme Buffer A, 10X: Supplied as a liquid in 33mM Tris-acetate, 10mM magnesium acetate, 66mM potassium acetate, 0.1mg/ml BSA (pH 7.9 at 37C). R1625-20: Restriction Enzyme Buffer for Bful, 10X: Supplied as a liquid in 10mM Tris-HCl (pH 7.4 at 25C), 300mMNaCl, 1mM DTT, 1mM EDTA, 0.2mg/ml BSA, 50% glycerol. Storage and Dilution Buffer: 10mM Tris-HCl (pH 7.4 at 25C), 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA, 50% glycerol. Thermal Inactivation: Bful is inactivated by incubation at 80C for 20min.