Protocol

Isolation of XO Subclones from XY Murine Embryonic Stem Cells

  1. Richard R. Behringer2
  1. 1Department of Genetics and Development, Columbia University Medical Center, New York, New York 10032, USA
  2. 2Department of Genetics, University of Texas MD Anderson Cancer Center, Houston, Texas 77030, USA
  1. 3Correspondence: vep1{at}columbia.edu

Abstract

This is a simple procedure to isolate XO subclones from XY murine embryonic stem cells in situations that require transmission of a mutation through the female germline—for example, if the mutation adversely affects spermatogenesis. XY cells are plated at clonal density, and resulting colonies are genotyped by polymerase chain reaction for a Y-specific probe to identify clones that have spontaneously lost the Y chromosome.

Footnotes

  • From the Mouse Phenotypes collection by Virginia E. Papaioannou and Richard R. Behringer.

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  1. Cold Spring Harb Protoc © 2023 Cold Spring Harbor Laboratory Press
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