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RELATED INFORMATION FAQs for NdeI FAQs for Restriction Endonucleases Technical Reference for Restriction Endonucleases High Fidelity (HF) Restriction Enzymes FAVORITE TOOLS Enzyme Finder NEBcutter NEBuffer Chart Double Digest Finder Isoschizomers DNA Sequences and Maps REBASE RELATED PRODUCTS Reagents Sold Separately NEBuffer 4

Home > Products > Restriction Endonucleases > Restriction Endonucleases > NdeI NdeI Catalog # Size Concentration Price Qty   R0111L 20,000 units 20,000 units/ml $232.00 R0111S 4,000 units 20,000 units/ml $58.00 Prices are in US dollars and valid only for US orders. Download: MSDS PDF Recognition Site: isoschizomers | compatible ends | single letter code Source: A E. coli strain that carries the NdeI gene from Neisseria denitrificans (NRCC 31009). Reagents Supplied: NEBuffer 4 Enzyme Properties Activity in NEBuffers: NEBuffer 1: 75% 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: (+ + +) Suitable for an extended or overnight digestion. Enzyme is active > 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: 20,000 units/ml Unit Assay Substrate: λ DNA Storage Conditions: 10 mM Tris-HCl 100 mM NaCl 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 A Notes General notes: DNA purified by standard miniprep procedures is cleaved at lower rates. FAQs Is NdeI a Time-Saver Qualified enzyme? Is NdeI activity sensitive to any type of substrate methylation? How many base pairs should be added at the end of a PCR primer after the NdeI recognition site to guarantee that NdeI will cut properly? Does NdeI have any isoschizomers? It seems that NdeI is having some difficulties cutting my DNA. Is there a reason for that? NdeI seems to be digesting my DNA correctly, but when I try to ligate it, I obtain no colonies. Is there a potential explanation for what I am observing? 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 20-fold overdigestion with NdeI, > 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 NdeI. 16-Hour Incubation: A 50 μl reaction containing 1 μg of DNA and 75 units of NdeI 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 4,000 units of NdeI with 1 μg of a mixture of single and double-stranded [3H] E. coli DNA (105 cpm/μg) for 4 hours at 37ºC released < 0.1% of the total radioactivity. Endonuclease Activity: Incubation of a 50 μl reaction containing 30 units of NdeI with 1 μg of ΦX174 RF I DNA for 4 hours at 37ºC resulted in < 15% conversion to RFII as determined by agarose gel electrophoresis. NdeI crystals (Lydia Dorner and Ira Schildkraut, New England Biolabs) Reagents Sold Separately NEBuffer 4