MssI (PmeI)

Produktübersicht

• Artikelname: MssI (PmeI)
• Artikelnummer: USB-M4692-60
• Hersteller Artikelnummer: M4692-60
• Alternativnummer: USB-M4692-60-250,USB-M4692-60-1250
• Hersteller: US Biological
• Kategorie: Molekularbiologie

Produktbeschreibung

5-G T T T A A A C-3 3-C A A A T T T G-5 Concentration: 5u/ul Source: Methylobacterium species Dd 5-732 Unit Definition: One unit is defined as the amount of Mssl required to digest 1ug of lambda DNA in 1 hour at 37C in 50ul of assay buffer. Diluent Buffer : 10mM Tris-HCl, pH 7.4, 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA and 50% glycerol. Storage Buffer: 10mM Tris-HCl, pH 7.5, 50mM KCl, 1mM DTT, 0.1mM EDTA, 0.2mg/ml BSA and 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 at 37C), 10mM magnesium acetate, 66mM potassium acetate, 0.1mg/ml BSA. R1625-04 Restriction Enzyme Buffer E, 10X: Dilute to 1X for 100% Mssl digestion. 10mM Tris-HCl, pH 7.5, 10mM MgCl2 and 0.1mg/ml BSA. Incubation Temperature: 37C Overdigestion Assay: No detectable change in the specific fragmentation pattern is observed after 160-fold overdigestion (10u/ug lambda DNA x 16 hours) with Mssl. Ligation/Recutting Assay: After 50-fold overdigestion (0.6u/ug DNA x 17 hours) with Mssl, more than 90% of the DNA fragments can be ligated in a mixture containing 20-40u of T4 ligase/ 1ug of fragments and 10% PEG at a 5-termini concentration of 0.09uM. 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 Mssl for 4 hours. Blue/White Cloning Assay: A mixture of pUC57/Hindlll, pUC57/Pstl and pUC57/Eco321 digests was incubated with 10 units of Mssl for 16 hours. After reeligation and transformation, the background level of white colonies was <1%. Thermal Inactivation: Mssl is inactivated by incubation at 65C for 20min. Stability during Prolonged Incubation: A minimum of 5 units of Mssl is required for complete digestion of 1ug of lambda DNA in 16 hours at 37C. Number of Recognition Sites in DNA: Lambda: 2 PhiX174: 0 M13mp18/19: 0 pBR322: 0 pUC18/19: 0 pUC57: 0 pTZ19R/U: 0 Protocol for Digestion: Add: Nuclease free water: 16ul R1625-04: 2ul DNA (0.5-1ug/ml): 1ul M4692-60: 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 R1625-04: 2ul M4692-60: 1-2ul Mix gently and spin down for a few seconds. Incubate at 37C for 1-16 hours. Storage and Stability: Aliquot and store at -20C. Aliquots are stable for at least 6 months at -20C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.