Exonuclease I

Exonuclease I (ExoI) degrades single-stranded DNA in a 3'→5' direction, releasing deoxyribonucleoside 5'-monophosphates in a  stepwise manner and leaving 5'-terminal dinucleotides intact. Detailní informace

Cena s DPH € 90.26
Cena bez DPH € 74.59
 4000 Units
Dostupnost Skladem
Kód produktu EN0581

Detailní popis Exonuclease I

Thermo Scientific Exonuclease I (ExoI) degrades single-stranded DNA in a  3'=>5' direction. It releases deoxyribonucleoside 5'-monophosphates in a stepwise manner and leaves 5'-terminal dinucleotides intact.

It does not cleave DNA strands with terminal 3'-OH groups blocked by phosphoryl or acetyl groups (see Reference 1).

Highlights

  • Active in PCR buffers

Applications

  • Primer removal from PCR mixtures:
    • prior to PCR product sequencing (see Reference 2)
    • for one-tube "megaprimer" PCR mutagenesis (see​ Reference 3)
  • Removal of single-stranded DNA containing a 3'-hydroxyl terminus from nucleic acid mixtures
  • Assay for the presence of single-stranded DNA with a 3'-hydroxyl terminus (see​ Reference 4)

Note

The enzyme is not suitable for removing 3'-overhangs of dsDNA.

10X Reaction Buffer 670 mM glycine-KOH (pH 9.5 at 25°C), 67 mM MgCl2, 10 mM DTT.
Definition of Activity Unit
  • One unit of the enzyme catalyzes the release of 10 nmol of acid soluble nucleotides in 30 min at 37°C.
  • Enzyme activity is assayed in the following mixture: 67 mM glycine-KOH (pH 9.5), 6.7 mM MgCl2, 1 mM DTT, and 0.17 mg/mL single-stranded [3H]-DNA.
Hazardous No
Inactivation Inactivated by heating in boiling water bath for 10 min, preferably in the presence of EDTA.
Inhibition Inhibitors: no specific inhibitors have been described.
Molecular Weight 54.5 kDa monomer.
Quality Control The absence of other endo-, exodeoxyribonucleases, and ribonucleases confirmed by appropriate quality tests.
Source E. coli cells with a cloned E. coli sbcB gene.
Storage Buffer The enzyme is supplied in:
20 mM Tris-HCl (pH 7.5), 0.1 mM EDTA, 1 mM DTT and 50% (v/v) glycerol.
Storage Condition -20 C

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