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One-hour universal protocol for mouse genotyping.

Romain Joubert1, Virginie Mariot1, Julie Dumonceaux1

  • 1NIHR Biomedical Research Centre, University College London, Great Ormond Street Institute of Child Health and Great Ormond Street Hospital NHS Trust, London, United Kingdom.

Muscle & Nerve
|February 23, 2020
PubMed
Summary

A new 1-hour universal genotyping (1-HUG) protocol simplifies the genotyping of genetically modified mice. This method is robust, cost-effective, and applicable to various mouse models and genomic modifications.

Keywords:
DMDFSHDgenotypingmouse modelneuromuscular diseases

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Area of Science:

  • Genetics
  • Molecular Biology
  • Animal Models

Background:

  • Genotyping is essential for research involving transgenic animals.
  • Existing protocols can be costly, time-consuming, and require specialized equipment.
  • Multiple protocols may be needed for different mouse strains, increasing complexity.

Purpose of the Study:

  • To develop a rapid and universally applicable genotyping protocol for genetically modified mice.
  • To overcome the limitations of existing genotyping methods in terms of time, cost, and versatility.

Main Methods:

  • Adaptation of the amplification-resistant mutation protocol.
  • Development of the 1-hour universal genotyping (1-HUG) protocol.
  • Streamlined workflow from sample isolation to PCR gel analysis.

Main Results:

  • The 1-HUG protocol successfully genotypes genetically modified mice in 1 hour.
  • Applicable to various mouse models, including mdx, FLExDUX4, and HSA-MerCreMer.
  • Effective for different genomic modifications and sex determination.

Conclusions:

  • The 1-HUG protocol offers a robust, efficient, and versatile solution for mouse genotyping.
  • Suitable for routine use in laboratories studying mouse models, particularly for neuromuscular diseases.