PaeI (SphI)

Produktübersicht

• Artikelname: PaeI (SphI)
• Artikelnummer: USB-P1800
• Hersteller Artikelnummer: P1800
• Alternativnummer: USB-P1800-500,USB-P1800-2500
• Hersteller: US Biological
• Kategorie: Molekularbiologie

Produktbeschreibung

Sequence: 5-G C A T G C-3 3-C G T A C G-5 Source: Pseudomonas aeruginosa Concentration: 10u/ul Unit Definition: One unit is defined as the amount of PaeI required to digest 1ug of lambda DNA fragments in 1 hr at 37C in 50ul assay buffer. Diluent Buffer: Supplied as a liquid in 10mM Tris-HCl, pH 7.4, 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA, 50% glycerol. Storage Buffer: 10mM potassium phosphate (pH 7.0 at 25C), 50mM KCl, 1mM DTT, 0.1mM EDTA, 0.5mg/ml BSA, 0.15% Triton X-100 and 50% glycerol. Supplied with: R1625: Supplied as a liquid in 33mM Tris-acetate, 10mM magnesium acetate, 66mM potassium acetate, 0.1mg/ml BSA, pH 7.9 at 37C. R1625-01: Restriction Enzyme Buffer B, 10X: Supplied as a liquid 10mM Tris-HCl, pH 7.5 at 37C, 10mM magnesium chloride, 0.1mg/ml BSA. Incubate at 37C Double Digests: R1625 is provided to simplify buffer selection for double digests. 98% of PaeI, works well in a 1x or 2x. Overdigestion Assay: No detectable change in the specific fragmentation pattern is observed after 160-fold overdigestion (10u/ug lambda DNA x 16 hours) with PaeI. Ligation Recutting Assay: After 50-fold overdigestion (3u/ug DNA x 17 hours) with PaeI, 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 10 units of PaeI for 4 hours. Blue White Cloning Assay: pUC57 was incubated with 10 units of PaeI for 16 hours. After religation and transformation <1% of white colonies were detected. Methylation Effects: Dam:, Dcm, EcoKI: never overlaps-no effect CpG: May overlap- no effect EcoBI: may overlap- blocked Stability during Prolonged Incubation: A minimum of 0.1 units of PaeI is required for complete digestion of 1ug of lambda DNA in 16 hours at 37C. Thermal Inactivation: PaeI is inactivated by incubation at 65C for 20min. Digestion of Agarose-embedded DNA: A minimum of 5 units of PaeI is required for digestion of 1ug of agarose-embedded lambda DNA in 16 hours. Number of Recognition Sites in DNA: Lambda 6 pBR322 1 phiX174 0 pUC57, pUC18/19, pTZ19R/U, M13mp18/19