Alw44I (ApaLI)

Produktübersicht

• Artikelname: Alw44I (ApaLI)
• Artikelnummer: USB-A1372-15
• Hersteller Artikelnummer: A1372-15
• Alternativnummer: USB-A1372-15-1000
• Hersteller: US Biological
• Kategorie: Molekularbiologie

Produktbeschreibung

5-G T G C A C-3 3-C A C G T G-5 Unit Definition: One unit is defined as the amount of Alw44l required to digest 1ug of lambda DNA-Smal fragments 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, 50% glycerol. Storage Buffer: Supplied as a liquid in 10mM Tris-HCl pH 7.5, 50mM KCl, 1mM DTT, 0.1mM EDTA, 0.2mg/ml BSA, 50% glycerol. Supplied with: R1625-Restriction Enzyme Buffer A (double digests): 1X Buffer composition-Supplied as a liquid containing 33mM Tris-acetate pH 7.9, 10mM magnesium acetate, 66mM potassium acetate, 0.1mg/ml BSA. Incubate at 37C. Methylation Effects on Digestion: Dam: never overlaps - no effect. Dcm: never overlaps - no effect. CpG: completely overlaps - blocked EcoKl: never overlap - no effect EcoBl: never overlaps - no effect. Number of Recognition Sites in DNA: Lambda: 4 PhiX174: 1 pBR322: 3 pUC57: 3 pUC18/19: 3 pTZ19R/U: 2 M13/mp18/19: 1 Compatible Ends: Bfml Digestion of Agarose-embedded DNA: A minimum of 5 units of the enzyme is required for complete digestion of 1ug of agarose-embedded lambda DNA in 16 hours. Overdigestion Assay: No detectable change in the specific fragmentation pattern is obseAlw44l.. rved after 160-fold overdigestion (10u/ug lambda DNA x 16 hours) with Alw44I Stability during Prolonged Incubation: A minimum of 0.1 units of the Alw44I is required for complete digestion of 1ug of DNA in 16 hours at 37C. Protocol for Digestion: Add: Nuclease free water: 16ul R1625: 2ul DNA (0.5-1ug/ml): 1ul A1372-15: 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 A1372-15: 1-2ul Mix gently and spin down for a few seconds. Incubate at 37C for 1-16 hours. Ligation/Recutting Assay: After 50-fold overdigestion (3u/ug DNA x 17 hours) with Alw44I more than 90% of the DNA fragments can be ligated at a 5-termini concentration of 0.03uM. More than 95% of these can be recut. Labeled Oligonucleotide (LO) Assay: No detectable degradation of a single-stranded and double-stranded labeled oligonucleotide occurred during incubation with 10 units of Alw44l for 4 hours. Thermal Inactivation: Enzyme is inactivated by incubation at 65C for 20 minutes.