Exonuclease III (E. coli)
- Double-stranded DNA specific exonuclease
- Initiates at the 3' termini of linear double-stranded DNA with 5' overhangs or blunt ends and 3' overhangs containing less than four bases
- Initiates at nicked sites in double-stranded DNA
- Catalyzes the removal of nucleotides from linear or nicked double-stranded DNA in the 3' to 5' direction
Exonuclease III is ideal for:
- Site-directed mutagenesis
- Preparation of single-stranded DNA for dideoxy sequencing
- Preparation of nested deletions in double-stranded DNA
Catalyzes the stepwise removal of mononucleotides from 3´-hydroxyl termini of duplex DNA (1). A limited number of nucleotides are removed during each binding event, resulting in coordinated progressive deletions within the population of DNA molecules (2).
The preferred substrates are blunt or recessed 3´-termini, although the enzyme also acts at nicks in duplex DNA to produce single-strand gaps. 3´-protruding termini are resistant to cleavage; the degree of resistance depends on the length of the extension,with extensions 4 bases or longer being essentially resistant to cleavage.
Exonuclease III activity depends partially on helical structure (4) and displays sequence dependence (C>A=T>G; ref. 5). Temperature, salt concentration and the ratio of enzyme to DNA greatly affect enzyme activity, requiring reaction conditions to be tailored to specific applications.
Exonuclease III has also been reported to have RNase H, 3´-phosphatase and AP-endonuclease activities (1).
Product SourcePurified from E.coli K-12, BE257/pSGR3 strain (kindly supplied by B. Weiss)
The following reagents are supplied with this product:
Store at (°C) Concentration NEBuffer™ 1 -20 10X
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