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FAQs for Cac8I FAQs for Restriction Endonucleases Technical Reference for Restriction Endonucleases Enzyme Finder NEBcutter NEBuffer Chart Double Digest Finder Isoschizomers DNA Sequences and Maps REBASE NEBuffer 4

Home > Products > Restriction Endonucleases > Restriction Endonucleases > Cac8I Cac8I Now NEBuffer 4 Catalog # Size Concentration Price Qty   R0579L 500 units 4,000 units/ml $264.00 R0579S 100 units 4,000 units/ml $66.00 Prices are in US dollars and valid only for US orders. Download: MSDS PDF Recognition Site: isoschizomers | compatible ends | single letter code Source: Clostridium acetobutylicum ABKn8 (G. Reysett) Reagents Supplied: NEBuffer 4 (10X) Enzyme Properties Activity in NEBuffers: NEBuffer 1: 50% NEBuffer 2: 75% NEBuffer 3: 100% 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: Blocked by some combinations of overlapping Heat Inactivation: 65°C for 20 minutes Survival in a Reaction: Minimum units to digest 1 µg of substrate DNA in 16 hours: 0.25 unit(s) 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 λ DNA in 1 hour at 37°C in a total reaction volume of 50 µl. Concentration: 4,000 units/ml Unit Assay Substrate: λ DNA Storage Conditions: 10 mM Tris-HCl 150 mM KCl 1 mM Dithiothreitol 0.1 mM EDTA 50% Glycerol 0.1% Triton X-100 pH 7.5 @ 25°C Storage Temperature: -20°C Diluent Compatibility: Diluent B FAQs The NEB catalog has historically stated that activity in NEBuffer4 is less than 100% yet this enzyme is now supplied with NEBuffer4. What have you changed? Has the conversion to NEBuffer4 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? Do degenerate recognition sites need to be palindromic? 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 5-fold overdigestion with Cac8I, > 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 Cac8I. 16-Hour Incubation: A 50 μl reaction containing 1 μg of DNA and 50 units of Cac8I 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 60 units of Cac8I with 1 μg of a mixture of single and double-stranded [3H] E. coli DNA (205 cpm/μg) for 4 hours at 37ºC released < 0.12% of the total radioactivity. Reagents Sold Separately NEBuffer 4