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FAQs for AluI FAQs for Restriction Endonucleases Technical Reference for Restriction Endonucleases Enzyme Finder NEBcutter NEBuffer Chart Double Digest Finder PCR Selection Tool Isoschizomers DNA Sequences and Maps REBASE NEBuffer 4

Home > Products > Restriction Endonucleases > Restriction Endonucleases > AluI AluI Catalog # Size Concentration Price Qty   R0137L 5,000 units 10,000 units/ml $244.00 R0137S 1,000 units 10,000 units/ml $61.00 Prices are in US dollars and valid only for US orders. Download: MSDS PDF Recognition Site: isoschizomers | compatible ends | single letter code Source: An E. coli strain that carries the cloned AluI gene from Arthrobacter luteus (ATCC 21606). Reagents Supplied: NEBuffer 4 (10X) Enzyme Properties Activity in NEBuffers: NEBuffer 1: 100% NEBuffer 2: 100% NEBuffer 3: 75% NEBuffer 4: 100% When using a buffer other than the optimal (supplied) NEBuffer, it may be necessary to add more enzyme to achieve complete digestion. Methylation Sensitivity: dam methylation: Not sensitive dcm methylation: Not sensitive CpG methylation: Not sensitive More information about: Methylation Sensitivity Heat Inactivation: 65°C for 20 minutes Survival in a Reaction: (+ +) Intermediate activity. Suitable for extended digestion, but < 8 hours. More information about: Extended Digests with Restriction Enzymes Reaction & Storage Conditions Reaction Conditions: 1X NEBuffer 4 Incubate at 37°C. 1X NEBuffer 4: 20 mM Tris-acetate 50 mM potassium acetate 10 mM Magnesium Acetate 1 mM Dithiothreitol pH 7.9 @ 25°C Unit Definition: One unit is defined as the amount of enzyme required to digest 1 µg of λ DNA in 1 hour at 37°C in a total reaction volume of 50 µl. Concentration: 10,000 units/ml Unit Assay Substrate: λ DNA Storage Conditions: 10 mM Tris-HCl 100 mM KCl 1 mM Dithiothreitol 0.1 mM EDTA 200 µg/ml BSA 50% Glycerol pH 7.4 @ 25°C Storage Temperature: -20°C Diluent Compatibility: Diluent B FAQs What is the activity of AluI at 25°C? Does AluI cleave ssDNA? The NEB catalog has historically stated that activity in NEBuffer 4 is less than 100% yet this enzyme is now supplied with NEBuffer 4. What have you changed? Has the conversion to NEBuffer 4 altered any of the properties of the restriction enzyme? If I have an old tube of enzyme, what NEBuffer should I use? Will the new enzyme work in the originally supplied NEBuffer? Why is NEB switching this restriction enzyme to NEBuffer 4? Are there any common problems with AluI? What is the molecular weight of AluI? Does a certain salt or salt level inhibit activity of AluI? Quality Control for Current Lot Quality control values for a specific lot can be found on the datacard which accompanies each product. Ligation and Re-cutting: After a 10-fold overdigestion with AluI, > 95% of the DNA fragments can be ligated with T4 DNA Ligase (at a 5' termini concentration of 1-2 μM) at 16ºC. Of these ligated fragments, > 95% can be recut with AluI. 16-Hour Incubation: A 50 μl reaction containing 1 μg of DNA and 100 units of AluI incubated for 16 hours at 37ºC resulted in a DNA pattern free of detectable nuclease degradation as determined by agarose gel electrophoresis. Exonuclease Activity: Incubation of a 50 μl reaction containing 100 units of AluI with 1 μg of a mixture of single and double-stranded [3H] E. coli DNA (205 cpm/μg) for 4 hours at 37ºC released < 0.1% of the total radioactivity. Reagents Sold Separately NEBuffer 4 Legal Patents: Invitrogen: Licensed Under U.S. Patent No. 5,334,526 New England Biolabs, Inc. is an ISO 9001 and ISO 14001 Certified Company. NEB certifies that it is a small business in accordance with the US Small Business Administration and 13 CFR 121.201