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FAQs for HpaII 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 > HpaII Methyltransferase HpaII Methyltransferase Catalog # Size Concentration Price Qty   M0214L 500 units 4,000 units/ml $224.00 M0214S 100 units 4,000 units/ml $56.00 Prices are in US dollars and valid only for US orders. Download: MSDS PDF Methylation Site: Description: HpaII Methyltransferase recognizes the same sequence as the MspI Methyltransferase, but modifies the internal cytosine residue (C5) of the sequence CCGG. Source: A E. coli strain that carries the cloned HpaII modification gene from Haemophilus parainfluenzae (ATCC 49669) Reagents Supplied: HpaII Methylase Reaction Buffer (10X) S-adenosylmethionine (SAM) Enzyme Properties Molecular Weight: Theoretical: 40,397 daltons Reaction & Storage Conditions Reaction Conditions: 1X HpaII Methylase Reaction Buffer Supplemented with 80 μM S-adenosylmethionine (SAM) Incubate at 37°C. 1X HpaII 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 HpaII restriction endonuclease. Protection Assay Conditions: HpaII Methyltransferase is incubated with 1 µg of λ DNA in 10 µl 1X HpaII 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 HpaII Methyltransferase is determined by the addition of 40 µl NEBuffer 1 supplemented with 10 mM MgCl2 and 10 units of HpaII restriction endonuclease. Incubation at 37°C for 30 minutes is followed by analysis on agarose gels. Concentration: 4,000 units/ml Storage Conditions: 50 mM Tris-HCl 100 mM NaCl 1 mM Dithiothreitol 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? Can HpaII Methyltransferase be heat inactivated? Does HpaII Methyltransferase require MgCl2? What is the molecular weight of HpaII Methyltransferase? Can HpaII methylated DNA be used to transform E. coli? Quality Control for Current Lot Quality control values for a specific lot can be found on the datacard which accompanies each product. Other Methylase Activity:: Incubation of 100 units of HpaII Methyltransferase with 1 µg λ DNA for 4 hours under standard Protection Assay Conditions did not protect the DNA from digestion by HpaI or MboI endonuclease. 16-Hour Incubation:: Incubation of 200 units of HpaII Methyltransferase with 1 μg of λ DNA in 50 μl 1X NEBuffer 2 for 16 hours at 37°C resulted in no detectable endonuclease contamination. Exonuclease Activity:: Incubation of 150 units of HpaII Methyltransferase with 1 μg sonicated 3H DNA (105 cpm/μg) for 4 hours at 37°C in 50 μl NEBuffer 2 released < 0.1% of the total radioactivity. References Hoffman, J.L. (1986) Biochemistry, 25, 4444-4449. Reagents Sold Separately S-adenosylmethionine (SAM)