User Manual
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1. lt gt MCLAB Molecular Cloning Laboratories User Manual Version 1 0 Revision Date 08 30 2013 Product name Exonuclease III E coli Cate Elll 100 Elll 200 EIll 300 EIII DOEM Description Exonuclease IlI E coli is a 3 gt 5 exonuclease which acts by digesting one strand of a dsDNA duplex at a time or digesting the RNA strand of an RNA DNA heteroduplex Exonuclease Ill E coli breaks phosphodi ester bonds on the 5 side of AP sites in both dsDNA and ssDNA It removes 3 terminal groups on dsDNA increases MutY turnover and efficiently degrades 3 recessed but not 3 protruding DNA ends creating 5 overhangs Exonuclease IIl E coli removes a limited number of nucleotides per binding event resulting in coordinated progressive deletions within the population of DNA molecules Specific Activity 150 000 U mg Application Degrades excess single stranded primer oligonucleotide from a reaction mixture containing double stranded extension products Source Purified from a strain of E coli that expresses the recombinant Exonuclease Ill gene Supplied in 25 mM Tris HCL 50 mM KCI 1 0 mM DTT 0 196 MM EDTA 5096 Glycerol pH 8 0 25 C Supplied with 10x Yellow Buffer 10x Yellow Buffer 100 mM Bis Tris Propane 100 mM MgCl 10 mM Dithiothreitol pH 7 0 25 C Unit Definition One unit is defined as the amount of enzyme required to produce 1 nmol of acid soluble total nucleotide i2. n 30 minutes at 37 C 1 650 872 0245 www mclab com 1 Recommended Storage Conditions 20 C Reference 1 Linn S M 1982 Nucleases pp 291 309 Cold Spring Harbor Laboratory Press 2 Shida T et al 1996 Nucl Acids Res 24 22 4572 4576 3 Doetsch P W 1990 Mutat Res 236 2 3 173 201 4 Pope M A et al 2002 J Biol Chem 277 25 22605 22615 5 Henikoff S 1984 Gene 28 351 359 2 S MCLAB
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