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Exonuclease III (E. coli )

No Product Name Cat.# Conc Size Price(₩) Add
1 Exonuclease III (E. coli ) M010S 100 units /μl 5,000 units ₩ 60,000 추가
2 Exonuclease III (E. coli ) M010L 100 units /μl 25,000 units ₩ 240,000 추가
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Product description

Exonuclease III (E.coli) is a 3’ → 5’ exonuclease specific for double-stranded DNA. The enzyme catalyzes the stepwise removal of mononucleotides starting from a 3’-OH at nicks, blunt ends, 5’ – overhangs and 3’ – overhangs of less than 4 bases, yielding nucleoside 5’ – phosphates.

A limited number of nucleotides are removed during each binding event, resulting in progressive deletions within the population of DNA molecules. Exonuclease III (E.coli) activity depends partially on helical structure and displays sequence dependence (C>A=T>G).Temperature, salt concentration and the ratio of enzyme to DNA greatly affect enzyme activity, requiring reaction conditions to be adjusted to specific applications.

 

Characteristics

- Molecular weight: 28 kDa

- Reaction temperature: 37

- Heat inactivation: 70 for 10 min

            

Applications

- Production of intermediates for site-directed mutagenesis

- Cloning of PCR products 

- Preparation of strand-specific probes

- Preparation of single-stranded templates for dideoxynucleotide sequencing

 

Quality control

- Purity: >99% on SDS-PAGE

- Endonuclease-free

            

Unit Definition

One unit is defined as the amount of enzyme required to produce 1nmol of acid-soluble nucleotides from double-stranded DNA in 30 minutes at 37.

 

Storage

5 mM KPO4, 200 mM KCl, 5 mM β-ME, 0.05 mM EDTA, 200 μg/ml BSA, 50% Glycerol, pH 6.5 @ 25

Store at -20.

 

10X Exonuclease III (E.coli) buffer

100 mM Bis-Tris-Propane-HCl, 100 mM MgCl2, 10 mM DTT pH 7.0

 

Cautions

- Exonuclease III is inhibited by Mg2+ chelator.

- Sulfhydryl compounds (DTT) stabilize the enzyme during long incubations.

- Exonuclease III is rapidly inactivated by heat in the absence of substrate DNA.

 

Standard reaction conditions

- Typical Reaction Conditions

10X Reaction Buffer

5 μl

DNA

2 μg

Exonuclease III

10 units

Sterile water

50 μl

→ incubate the reaction mixture at 37

 

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