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Exonuclease III (E. coli)

Catalog # Concentration Size List Price Quantity Your Price
M0206L 100000 units/ml 25000 units $409.00
$368.10
M0206S 100000 units/ml 5000 units $103.00
$92.70
Catalog # Size List Price Your Price
M0206L 25000 units $409.00
$368.10
M0206S 5000 units $103.00
$92.70
Catalog #
Qty:
 
*On-line ordering is for Canadian customers only. Web pricing is applicable only to orders placed online at www.neb.ca

  • 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
  • Need help finding the right exonuclease for your experiments? Try Exo Selector 

Exonuclease III is ideal for:

  • Site-directed mutagenesis
  • Preparation of single-stranded DNA for dideoxy sequencing
  • Preparation of nested deletions in double-stranded DNA
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  • Monarch_MiniprepProtocol_VideoThumb
    What are exonucleases and their applications?
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). Exonuclease III has also been reported to have RNase H, 3´-phosphatase and AP-endonuclease activities (1).

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. 
Product Source
An E. coli strain that carries the cloned Exonuclease III gene from E. coli.
Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  Exonuclease III (E. coli) M0206SVIAL -20 1 x 0.05 ml 100,000 units/ml
  NEBuffer™ 1 B7001SVIAL -20 1 x 1.25 ml 10 X
  Exonuclease III (E. coli) M0206LVIAL -20 1 x 0.25 ml 100,000 units/ml
  NEBuffer™ 1 B7001SVIAL -20 1 x 1.25 ml 10 X
Application Features
  • Unidirectional nested deletions (3)
  • Site-directed mutagenesis (6)
  • Preparation of strand-specific probes (2)
  • Preparation of single-stranded substrates for dideoxy sequencing (7)

Properties & Usage

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 [3H]-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

Companion Products
Materials Sold Separately
Notes
  • Phosphorothioate linkages are not cleaved by Exonuclease III. Unidirectional deletions can also be created by protecting one terminus by incorporation of α-phosphorothioate-containing nucleotide.

  • 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%
References
  • Rogers, G.S. and Weiss, B. (1980). L. Grossman and K. Moldave(Ed.), Methods Enzymol.. 65, 201-211. New York: Academic Press.
  • Sambrook, J., Fritsch, E.F. and Maniatis, T. (1989). Molecular Cloning: A Laboratory Manual (2nd. Ed.). 5.84-5.85.
  • Henikoff, S. (1984). Gene. 28, 351-359.
  • Richardson, C.C. et al. (1964). J. Biol. Chem. 239, 251-258.
  • Linxweiler, W. and Horz, W. (1982). Nucl. Acids Res. 10, 4845-4859.
  • Vandeyar, M.A. (1988). Gene. 65, 129-133.
  • Guo, L.H. and Wu, R. (1982). Nucl. Acids Res.. 10, 2065-2084.
  • Putney, S., Benkovic, S. and Schimmel, P. (1981). Proc. Natl. Acad. Sci. USA. 78, 7350-7354.
Protocols
Quality Control Assay
Quality Control tests are performed on each new lot of NEB product to meet the specifications designated for it. Specifications and individual lot data from the tests that are performed for this particular product can be found and downloaded on the Product Specification Sheet, Certificate of Analysis, data card or product manual. Further information regarding NEB product quality can be found here.
Specifications
The Specification sheet is a document that includes the storage temperature, shelf life and the specifications designated for the product. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]
Certificate of Analysis
The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality controls for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]
Safety Data Sheets
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