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Exonuclease III / 4KU / Cat.#3005
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  Exonuclease III

    

#3005

 

4,000 units

#3005L

 

20,000 units

 

 

DESCRIPTION 

Exonuclease III catalyzes the stepwise removal of 5' mononucleotides from the 3'-hydroxyl termini of double-stranded DNA.  Linear double-stranded DNA and circular DNAs containing nicks or gaps are substrates. The enzyme is not active on single-stranded DNA or double-stranded DNA with a protruding 3' terminus.  It will not degrade double-stranded DNA with 3' overhang of four bases or longer.  The enzyme also carries three other activities: an endonuclease specific for apurinic DNA, an RNase H activity, and a 3' phosphatase activity.  

 

 

SOURCE 

Recombinant E. coli strain. 

 

 

APPLICATIONS 

¡Ü Generating nested sets of deletions of the terminal 

sequences of double-stranded linear DNAs. 

¡Ü Site-directed mutagenesis. 

¡Ü Preparation of single-stranded substrates for 

   dideoxy sequencing.  

 

 

 

UNIT DEFINITION 

One unit is defined as the amount of enzyme required to produce 1nmole of acid-soluble nucleotides in 30 minutes at 37¡É. 

 

 

STANDARD UNIT ASSAY CONDITIONS 

50mM Tris-HCl (pH7.6), 1mM MgCl2, 1mM DTT and 125¥ìg calf thymus DNA. 

 

 

CONCENTRATION: 150-200 units/¥ìl. 

 

 

10X REACTION BUFFER 

500mM Tris-HCl (pH8.0@25¡É), 50mM MgCl2, 10mM 2-mercaptoethanol. 

 

 

STORAGE CONDITIONS 

20mM Tris-HCl (pH7.9), 1mM DTT. 160mM KCl and 50% glycerol.  Store at -20¡É.¡¡  

 

 

REFERENCES 

1. Rogers, S.G. and Weiss, B. (1980) Methods. Enzymol. 65, 201. 

2. Weiss, B. (1976) J. Biol. Chem. 251, 1896. 

3. Sambrook, J., Fritsch, E.F. and Maniatis, T. (1989) Molecular Cloning: A Laboratory Manual, second ed., Cold Spring Harbor Laboratory, Cold Spring Harbor. 

 


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