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FAQs for HhaI 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 > HhaI Methyltransferase HhaI Methyltransferase Catalog # Size Concentration Price Qty   M0217L 5,000 units 25,000 units/ml $244.00 M0217S 1,000 units 25,000 units/ml $61.00 Prices are in US dollars and valid only for US orders. Download: MSDS PDF Methylation Site: Description: HhaI Methyltransferase modifies the internal cytosine residue (C5) of the sequence GCGC. Source: A E. coli strain that carries the cloned HhaI modification gene from Haemophilus haemolyticus (ATCC 10014) Reagents Supplied: HhaI Methylase Reaction Buffer (10X) S-adenosylmethionine (SAM) Reaction & Storage Conditions Reaction Conditions: 1X HhaI Methylase Reaction Buffer Supplemented with 32 mM S-adenosylmethionine (SAM) Incubate at 37°C. 1X HhaI Methylase 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 λ DNA in 1 hour at 37°C in a total reaction volume of 10 µl against cleavage by HhaI restriction endonuclease. Protection Assay Conditions: HhaI Methyltransferase is incubated with 1 µg of λ DNA in 10 µl 1X HhaI 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 HhaI Methyltransferase is determined by the addition of 40 µl NEBuffer 4 supplemented with 100 µg/ml BSA, 10 mM MgCl2 and 10 units of HhaI restriction endonuclease. Incubation at 37°C for 30 minutes is followed by analysis on an agarose gel. Concentration: 25,000 units/ml Storage Conditions: 50 mM Tris-HCl 150 mM NaCl 5 mM 2-Mercaptoethanol 10 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. FAQs Is S-adenosylmethionine (SAM) supplied with the Methyltransferase? What should be considered if the methylation is not going to completion? What is the molecular weight of HhaI Methyltransferase? 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 125 units of HhaI Methyltransferase with 1 μg of HindIII-digested DNA in 50 μl of 1X NEBuffer 2 for 16 hours at 37°C resulted in no detectable endonuclease contamination. Exonuclease Activity:: Incubation of 250 units of HhaI Methyltransferase with 1 μg sonicated 3H DNA (105 cpm/μg) for 4 hours at 37°C in 50 μl NEBuffer 2 released 0.39% of the total radioactivity. M.HhaI crystal (Xiaodong Cheng, Cold Spring Harbor Laboratory) References Hoffman, J.L. (1986) Biochemistry, 25, 4444-4449. Reagents Sold Separately S-adenosylmethionine (SAM)