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RiboTag Immunoprecipitation in the Germ Cells of the Male Mouse
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Sex Genotyping Mice by Polymerase Chain Reaction.

Virginia E Papaioannou1, Richard R Behringer2

  • 1Department of Genetics and Development, Columbia University Medical Center, New York, New York 10032, USA vep1@columbia.edu.

Cold Spring Harbor Protocols
|November 6, 2023
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Summary

Determine mouse genetic sex using a simple polymerase chain reaction (PCR) method. This technique amplifies the male-specific Sry gene from tail or ear tissue, efficiently identifying the Y-chromosome.

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Area of Science:

  • Genetics
  • Molecular Biology
  • Animal Science

Background:

  • Accurate determination of genetic sex in laboratory mice is crucial for research.
  • Traditional methods can be invasive or time-consuming.
  • A need exists for a simple, reliable sexing protocol.

Purpose of the Study:

  • To present a straightforward polymerase chain reaction (PCR) protocol for determining mouse genetic sex.
  • To validate the detection of the Y-chromosome-specific Sry gene for sex determination.

Main Methods:

  • DNA extraction from mouse tissue samples (tail tip or ear punch).
  • Polymerase chain reaction (PCR) amplification targeting the male-specific Sry gene.
  • Detection of amplified Sry gene product to indicate male sex.

Main Results:

  • Successful amplification of the Sry gene was achieved from male mouse samples.
  • The PCR protocol demonstrated high specificity for the Y-chromosome.
  • Tissue lysates from tail tip and ear punch were suitable for DNA analysis.

Conclusions:

  • This PCR-based method provides a simple and effective means to determine the genetic sex of mice.
  • The Sry gene serves as a reliable molecular marker for identifying males.
  • The protocol is practical for routine use in laboratory animal facilities.