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Related Experiment Video

Updated: Aug 26, 2025

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Automated multi-sample DNA extraction for genotyping live Xenopus embryos.

Narmadha Alles1, Matthew Guille2, Dariusz C Górecki1

  • 1School of Pharmacy and Biomedical Sciences, University of Portsmouth, Portsmouth, UK.

Developmental Dynamics : an Official Publication of the American Association of Anatomists
|October 11, 2022
PubMed
Summary

Rapid DNA extraction from early-stage Xenopus embryos enables safe and ethical identification of genetic mutants. This method supports early analysis and reduces animal welfare concerns, aligning with the 3Rs principles.

Keywords:
CRISPR/Cas9XenopusZebrafish Embryonic Genotypergenotyping

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Area of Science:

  • Amphibian research
  • Developmental biology
  • Genetics

Background:

  • Xenopus frogs are valuable models for genetic disease research due to their reproductive capacity and rapid embryonic development.
  • Current Xenopus genotyping methods are inefficient, requiring numerous embryos or late-stage sampling with welfare implications.

Purpose of the Study:

  • To adapt the Zebrafish Embryonic Genotyper for early-stage genomic DNA extraction in Xenopus.
  • To enable rapid and non-invasive genotyping of Xenopus embryos.

Main Methods:

  • Genomic DNA extraction from Xenopus tropicalis and Xenopus laevis at early developmental stages (Nieuwkoop and Faber stage 19).
  • Adaptation of the Zebrafish Embryonic Genotyper for Xenopus DNA extraction.
  • Amplification of DNA fragments up to 800 bp for downstream analyses.

Main Results:

  • Sufficient, high-quality DNA was extracted from early-stage Xenopus embryos.
  • The DNA extraction process showed no adverse effects on tadpole development, behavior, or morphology.
  • Successful amplification and downstream analysis (gel electrophoresis, T7 endonuclease I assay, Sanger sequencing) of extracted DNA.

Conclusions:

  • The adapted method allows for rapid genotyping early in Xenopus development.
  • Early identification of mutants facilitates early analysis or selective breeding, improving resource efficiency.
  • The method aligns with the 3Rs principles (Replacement, Reduction, Refinement) by minimizing invasive procedures and improving animal welfare.