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FAQs for AluI Methyltransferase FAQs for DNA Modifying Enzymes and Cloning Technical Reference for DNA Modifying Enzymes and Cloning Enzyme Finder NEBcutter NEBuffer Chart Double Digest Finder Isoschizomers DNA Sequences and Maps REBASE S-adenosylmethionine (SAM)

Home > Products > DNA Modifying Enzymes and Cloning > Methyltransferases > AluI Methyltransferase AluI Methyltransferase Catalog # Size Concentration Price Qty   M0220L 500 units 5,000 units/ml $244.00 M0220S 100 units 5,000 units/ml $61.00 Prices are in US dollars and valid only for US orders. Download: MSDS PDF Methylation Site: Description: AluI Methyltransferase modifies the cytosine residue (C5) of the sequence AGCT. Source: A E. coli strain that carries the cloned AluI modification gene from Arthrobacter luteus (ATCC 21606). Reagents Supplied: AluI Methyltransferase Reaction Buffer (10X) S-adenosylmethionine (SAM) Reaction & Storage Conditions Reaction Conditions: 1X AluI Methyltransferase Reaction Buffer Supplemented with 32 μg/ml S-adenosylmethionine (SAM) Incubate at 37°C. 1X AluI Methyltransferase Reaction Buffer: 50 mM Tris-HCl 5 mM 2-Mercaptoethanol 10 mM EDTA pH 7.5 @ 25°C Unit Definition: One unit is defined as the amount of enzyme required to protect 1 µg of λ DNA in 1 hour at 37°C in a total reaction volume of 10 µl against cleavage by AluI restriction endonuclease. Protection Assay Conditions: AluI Methyltransferase is incubated with 1 µg of λ DNA in 10 µl 1X AluI Methyltransferase Buffer, supplemented with 80 µM S-adenosylmethionine, for one hour at 37°C followed by 15 minutes at 65°C. The extent of protection by AluI Methyltransferase is determined by  the addition of 40 µl NEBuffer 1 supplemented with 10 mM MgCl2 and 10 units of AluI restriction endonuclease. Incubation at 37°C for 30 minutes is followed by analysis on an agarose gel. Concentration: 5,000 units/ml Storage Conditions: 10 mM Tris-HCl 50 mM KCl 1 mM Dithiothreitol 0.1 mM EDTA 200 µg/ml BSA 50% Glycerol pH 7.5 @ 25°C Storage Temperature: -20°C Notes General notes: Storage of SAM: S-adenosylmethionine is stored at –20°C as 32 mM solution dissolved in sulfuric acid (0.005 M) and 10% ethanol. SAM in this solution stored under ideal conditions remains active for up to 6 months. SAM is unstable at (pH 7.5), 37°C, and should be replenished for reactions incubated longer than 4 hours. Many problems in achieving complete digestion can be alleviated by using fresh SAM. AluI methylated DNA is mcrB sensitive. FAQs Is S-adenosylmethionine (SAM) supplied with the Methyltransferase? What should be considered if the methylation is not going to completion? Can AluI Methyltransferase be heat inactivated? Does AluI Methyltransferase require MgCl2? What is the molecular weight of AluI Methyltransferase? Can AluI methylated DNA be used to transform E. coli? Does methylation with AluI Methyltransferase block SacI restriction endonuclease? Quality Control for Current Lot Quality control values for a specific lot can be found on the datacard which accompanies each product. 16-Hour Incubation:: Incubation of 40 units of AluI Methyltransferase with 1 μg λ DNA in 50 μl of 1X NEBuffer 2 at 37°C resulted in no detectable endonuclease contamination. Exonuclease Activity:: Incubation of 40 units of AluI Methyltransferase with 1 μg sonicated 3H DNA (105 cpm/μg) for 4 hours at 37°C in 50 μl NEBuffer 2 released < 0.2% of the total radioactivity. References Hoffman, J.L. (1986) Biochemistry, 25, 4444-4449. Reagents Sold Separately S-adenosylmethionine (SAM)