PdmI (XmnI)

Produktübersicht

• Artikelname: PdmI (XmnI)
• Artikelnummer: USB-P3125
• Hersteller Artikelnummer: P3125
• Alternativnummer: USB-P3125-500,USB-P3125-2500
• Hersteller: US Biological
• Kategorie: Molekularbiologie

Produktbeschreibung

5-G A A N NN N T T C-3 3-C T T N NN N A A G-5 Source: Pseudomonas diminuta Auk 5-324 Concentration: 10u/ul Unit Definition: One unit is defined as the amount of PdmI required to digest 1ug of lambda DNA in 1 hour at 37C in 50ul of assay buffer. Storage Buffer: 10mM Tris-HCl, pH 7.5, 100mM KCl, 1mM DTT, 5mM MgCl2, 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, 10mM magnesium acetate, 66mM potassium acetate, 0.1mg/ml BSA Incubation Temperature: 37C Thermal Inactivation: PdmI is inactivated by incubation at 65C for 20 minutes. Dilution Buffer: 10mM Tris-HCl, pH 7.4, 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA, 50% glycerol. Enzyme Properties: Methylation Effects on Digestion: Dam: Never overlaps - no effect Dcm: Never overlaps - no effect CpG: May overlap - cleavage impaired EcoKI: Never overlaps - effect not determined EcoBI: May overlap - effect not determined Stability during Prolonged Incubation: A minimum of 0.5 units of enzyme is required for complete digestion of 1ug of lambda DNA in 16 hours at 37C. Digestion of Agarose-embedded DNA: A minimum of 10 units of enzyme is required for complete digestion of 1ug of agarose-embedded lambda DNA in 16 hours. Number of Recognition Sites in DNA: Lambda: 24 PhiX174: 3 pBR322: 2 pUC57: 1 pUC18/19: 1 pTZ19R/U: 1 M13mp18/19: 2 Overdigestion Assay: No detectable change in the specific fragmentation pattern is observed after 160-fold overdigestion with Pdml (10u/ug lambda DNA x 16 hours). Ligation/Recutting Assay: The ligation and recleavage assay was replaced with L0 test after validating experiments showed L0 test ability to trace nuclease and phosphatase activities with sensitivity that is higher than L/R by a factor of 100. Labeled Oligonucleotide Assay: No detectable degradation of a single-stranded or double-stranded labeled oligonucleotide was observed after incubation with 10 units of Pdml for 4 hours. Protocol for Digestion: Add: Nuclease free water: 16ul R1625: 2ul DNA (0.5-1ug/ml): 1ul Pdml: 0.5-2ul Mix gently and spin down for a few seconds. Incubate at 37C for 1-2 hours. The digestion reaction may be scaled either up or down. 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: 2ul Pdml: 1-2ul Mix gently and spin down for a few seconds. Incubate at 37C for 1-16 hours. Storage and Stability: May be stored at 4C. For long-term storage, aliquot and store at -20C. Aliquots are stable for 6 months after receipt. For maximum recovery of product, centrifuge the original vial prior to removing the cap. Further dilutions can be made in assay buffer.