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Updated: Jul 7, 2026

Atomic Force Microscopy Investigations of DNA Lesion Recognition in Nucleotide Excision Repair
10:59

Atomic Force Microscopy Investigations of DNA Lesion Recognition in Nucleotide Excision Repair

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Endonucleolytic mutation analysis by internal labeling (EMAIL).

Michael J Cross1, Daniel L E Waters, L Slade Lee

  • 1Grain Foods CRC, Centre for Plant Conservation Genetics, Southern Cross University, Lismore, NSW, Australia. mcross@scu.edu.au

Electrophoresis
|February 22, 2008
PubMed
Summary
This summary is machine-generated.

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This study introduces Endonucleolytic Mutation Analysis by Internal Labeling (EMAIL), a novel method for detecting DNA variations. EMAIL improves sensitivity and reduces background noise compared to traditional methods, enabling clearer identification of genetic mutations.

Area of Science:

  • Molecular Biology
  • Genetics
  • Biotechnology

Background:

  • Mismatch-specific endonucleases are used for DNA variation scanning.
  • Conventional methods use 5'-labeled amplicons, facing limitations in signal strength and background noise.
  • Existing fluorescent and intercalating dye systems have similar pooling capacities for mismatch scanning.

Purpose of the Study:

  • To investigate CEL nuclease activity with internally and 5'-labeled amplicons.
  • To develop an improved method for mismatch detection with enhanced sensitivity and reduced background.
  • To introduce Endonucleolytic Mutation Analysis by Internal Labeling (EMAIL) for DNA variation analysis.

Main Methods:

  • Generated amplicons from rice starch synthase IIa gene (exon 8).

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Proofreading and DNA Repair Assay Using Single Nucleotide Extension and MALDI-TOF Mass Spectrometry Analysis
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Proofreading and DNA Repair Assay Using Single Nucleotide Extension and MALDI-TOF Mass Spectrometry Analysis

Published on: June 19, 2018

Related Experiment Videos

Last Updated: Jul 7, 2026

Atomic Force Microscopy Investigations of DNA Lesion Recognition in Nucleotide Excision Repair
10:59

Atomic Force Microscopy Investigations of DNA Lesion Recognition in Nucleotide Excision Repair

Published on: May 24, 2017

Comparative Lesions Analysis Through a Targeted Sequencing Approach
08:16

Comparative Lesions Analysis Through a Targeted Sequencing Approach

Published on: November 5, 2019

Proofreading and DNA Repair Assay Using Single Nucleotide Extension and MALDI-TOF Mass Spectrometry Analysis
11:08

Proofreading and DNA Repair Assay Using Single Nucleotide Extension and MALDI-TOF Mass Spectrometry Analysis

Published on: June 19, 2018

  • Labeled amplicons internally and at 5'-termini.
  • Investigated CEL nuclease activity on labeled substrates.
  • Developed and applied the EMAIL assay for mismatch detection.

    • Main Results:

    • Internal labeling significantly reduced background noise compared to 5'-labeling.
    • CEL nuclease activity reduced 5'-label signals but minimally affected internal labels.
    • 5'-modification enhanced exonucleolytic activity, removing terminal nucleotides.
    • The EMAIL assay correctly reclassified a rice line as heterozygous for a starch gene, contrary to previous findings.

    Conclusions:

    • Internal labeling offers improved sensitivity and reduced background for mismatch detection.
    • EMAIL provides a robust alternative to conventional methods for identifying subtle DNA variations.
    • EMAIL demonstrates potential for accurate identification of multiple mutations in allelic pools.