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A Fluorescence-based Exonuclease Assay to Characterize DmWRNexo, Orthologue of Human Progeroid WRN Exonuclease, and Its Application to Other Nucleases
JoVE Journal
Biologia
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JoVE Journal Biologia
A Fluorescence-based Exonuclease Assay to Characterize DmWRNexo, Orthologue of Human Progeroid WRN Exonuclease, and Its Application to Other Nucleases

A Fluorescence-based Exonuclease Assay to Characterize DmWRNexo, Orthologue of Human Progeroid WRN Exonuclease, and Its Application to Other Nucleases

DOI:
10.3791/50722-v

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06:10 min

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December 23, 2013

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, ,
1Department of Biochemistry,University of Oxford

Capitoli

  • 00:05Titolo
  • 00:54His-trap Purification
  • 02:24Nuclease Assay
  • 04:14Results: The Exonuclease Activity of DmWRNexo
  • 05:51Conclusion

Summary

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Exonucleases play critical roles in ensuring genome stability. Loss of WRN exonuclease function results in premature aging. Studying substrates and other requirements of the nuclease in vitro can help elucidate its role in vivo. Here we demonstrate a rapid and reproducible fluorescence-based assay to measure its nuclease activity.

Tags

Keywords: Fluorescence-based AssayDmWRNexoWRN ExonucleaseDNA DamageGenome StabilityNuclease ActivityDNA ReplicationRecombinationOligonucleotide SubstrateRadioactive AssayProcessivitySubstrate Preference

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