New England Biolabs, M0206L, Exonuclease III (E. coli)

Related Categories Exonucleases and Non-specific Endonucleases Applications Polymerases for DNA Manipulation Specification Unit Definition One unit is defined as the amount of enzyme required to produce 1 nmol of acid-soluble total nucleotide in a total reaction volume of 50 μl in 30 minutes at 37°C in 1X NEBuffer 1 with 0.15 mM sonicated duplex [ 3 H]-DNA. Reaction Conditions 1X NEBuffer™ 1 Incubate at 37°C 1X NEBuffer™ 1 10 mM Bis-Tris-Propane-HCl 10 mM MgCl2 1 mM DTT (pH 7 @ 25°C) Storage Buffer 5 mM KPO4 200 mM KCl 5 mM β-ME 0.05 mM EDTA 200 µg/ml BSA 50% Glycerol pH 6.5 @ 25°C Heat Inactivation 70°C for 20 minutes Specific Activity 150,000 units/mg FAQ Q: What is the activity of M0206 Exonuclease III in various NEB buffers? A: The activity of Exonuclease III in various buffers is listed below: NEBuffer 1 - 100% NEBuffer 2 - 75% NEBuffer 3 -25% NEBuffer 4 - 75% CutSmart – 100% Q: How does T5 Exonuclease differ from Exonuclease III (NEB# M0206)? A: Exonuclease III acts in the 3' to 5' direction, opposite that of T5 Exonuclease, which acts in the 5’ to 3’ direction. Q: How Does T7 Exonuclease differ from Exonuclease III (NEB# M0206)? A: Exonuclease III acts in the 3' to 5' direction, opposite that of T7 Exonuclease and Lambda Exonuclease. Q: Will Exonuclease III work in rCutSmart buffer? A: Exonuclease III will work in rCutSmart Buffer. To see its % functional activity in rCutSmart, and that of other DNA modifying enzymes in the cloning workflow, refer to the Activity of DNA Modifying Enzymes in rCutSmart® Buffer chart. Q: Can Exonuclease I be used with a double stranded exonuclease to clean up plasmid preparations? A: Exonuclease I can be used with Lambda Exonuclease (NEB# M0262) to clean up plasmid preps. Exonuclease III (NEB# M0206) and T7 Exonuclease (NEB# M0263) will also work, but will damage nicked plasmids. Although Exonuclease I can be used, we recommend using Exonuclease V (RecBCD) (NEB #M0345) to remove chromosomal DNA after plasmid prep (see our Application Note: Using Exonuclease V (RecBCD) to Eliminate Residual Genomic DNA When Purifying Low Copy Plasmids with the Monarch® Plasmid Miniprep Kit.