85-1378-83　AluI 600U　A1365

特徴

• 5'-A G^C T-3' 3'-T C^G A-5'
• Source:Arthrobacter luteus
• Concentration:10u/ul
• Supplied With:R1625 - Restriction Enzyme Buffer A, 10X: Dilute to 1X for use. 1X buffer composition is 33mM Tris-acetate (pH 7.9 at 37°C), 10mM magnesium acetate, 66mM potassium acetate, 0.1mg/ml BSA
• Diluent Buffer:10mM Tris-HCl (pH 7.4), 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA and 50% glycerol
• Unit Definition:One unit is defined as the amount of Alul required to digest 1ug of lambda DNA in 1 hour at 37°C in 50ul of assay buffer.
• Stability during Prolonged Incubation:A minimum of 0.1units of enzyme is required for complete digestion of 1ug of lambda DNA in 16 hours at 37°C.
• Thermal Inactivation:Enzyme is inactivated by incubation at 65°C for 20 minutes
• Methylation Effects:Dam, Dcm, CpG,EcoKl: never overlaps - no effect
• EcoBl: may overlap - blocked
• Number of Recognition Sites in DNA:Lambda: 143 PhiX174: 24 pBR322: 1pUC5: 16pUC18/19: 16pTZ19R/U: 1M13mp18/19: 27
• Overdigestion Assay: No detectable change in the specific fragmentation pattern is observed after 160-fold overdigestion (10u/ug lambda DNA x 16 hours) with AluI.
• Ligation/Recleavage Assay:The ligation and recleavage assay was replaced with LO test after validating experiments showed LO test ability totrace nuclease and phosphatase activities with sensitivity that is higher than L/R by a factor of 100.
• Labeled Oligonucleotide (LO) Assay:No detectable degradation of a single-stranded and double-stranded labeled oligonucleotide was observed after incubation with 10units of restriction endonuclease for 4 hours.
• Protocol for Digestion:Add:Nuclease free water: 16ulR1625: 2ulDNA (0.5-1ug/ml): 1ulA1365: 0.5-2ulMix gently and spin down for a few seconds. Incubate at 37ºC for 1-16 hours.
• Protocol for Digestion Directly after Amplification:Add:PCR Reaction Mixture: 10ul (~0.1-0.5ug of DNA)Nuclease free water: 18ulR1625: 2ulA1365: 1-2ulMix gently and spin down for a few seconds. Incubate at 37ºC for 1-16 hours.