Alw26I (BsmAI)

Produktübersicht

• Artikelname: Alw26I (BsmAI)
• Artikelnummer: USB-A1372-10
• Hersteller Artikelnummer: A1372-10
• Alternativnummer: USB-A1372-10-1000
• Hersteller: US Biological
• Kategorie: Molekularbiologie

Produktbeschreibung

5-G T C T C (N)1 -3 3-C A G A G (N)5 -5 Source: Acinetobacter lwoffi RFL26 Concentration: 10u/ul Unit Definition: One unit is defined as the amount of Alw26I required to digest 1ug of lambda DNA in 1 hour at 37C in 50ul of assay buffer. Incubation Temperature: 37C Diluent Buffer: 10mM Tris-HCl, pH 7.4, 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA and 50% glycerol. Storage Buffer: Supplied as a liquid in 10mM Tris-HCl, pH 7.4, 100mM KCl, 1mM DTT, 1mM EDTA, 0.2mg/ml BSA and 50% glycerol. Restriction Enzyme Buffer A, 10X: R1625: Supplied as a liquid in 33mM Tris-acetate, pH 7.9, 10mM magnesium acetate, 66mM potassium acetate and 0.1mg/ml BSA. Incubate at 37C. Supplied to simplify buffer selection for double digests. 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 Alw26I. Ligation/Recutting Assay: After 50-fold overdigestion (3u/ug DNA x 17 hours) with Alw26I more than 95% of the DNA fragments can be ligated at a 5-termini concentration of 0.1uM. 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 Alw26I for 4 hours. Methylation Effects: Alw26I does not cut GAGAm5C. Overlapping CpG methylation may influence DNA cleavage. Stability during Prolonged Incubation: A minimum of 0.2units of Alw26I is required for complete digestion of 1ug of lambda DNA in 16 hours at 37C. Thermal Inactivation: Alw26I is inactivated by incubation at 65C for 20min. Number of Recognition Sites in DNA: Lambda: 37 PhiX174: 4 M13mp18/19: 5 pBR322: 3 pUC18/19: 4 pUC57: 4 pTZ19R/U: 2 Protocol for Digestion: Add: Nuclease free water: 16ul R1625: 2ul DNA (0.5-1ug/ml): 1ul A1372-10: 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: 2ul A1372-10: 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 short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20C. Aliquots are stable for 6 months after receipt 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.