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Isolation and Analysis of the CGG-Repeat Size in Male and Female Gametes from a Fragile X Mouse Model

  • Xiaonan Zhao
  • Huiyan Lu
  • Pradeep K. Dagur
  • Karen UsdinEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 2056)

Abstract

Analysis of individual gametes has a number of applications in the study of the mechanism of repeat expansion in mouse models of the fragile X-related disorders, as well as in mouse models of other repeat expansion diseases. This chapter describes the techniques required to isolate oocytes and male gametes of different stages of maturity, along with the techniques required to accurately determine the repeat number in these gametes.

Keywords

Oocyte Sperm Germ cell Fluorescence-activated cell sorting Single cell analysis Small pool PCR 

Notes

Acknowledgments

The authors would like to thank Dr. John Philip McCoy, Jr. (NHLBI) for his careful reading of the manuscript. This work was made possible with funding to KU from the Intramural Program of NIDDK, NIH (DK057808).

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  • Xiaonan Zhao
    • 1
  • Huiyan Lu
    • 2
  • Pradeep K. Dagur
    • 3
  • Karen Usdin
    • 1
    Email author
  1. 1.Section on Gene Structure and Disease, Laboratory of Cell and Molecular Biology, National Institute of Diabetes, Digestive and Kidney DiseasesNational Institutes of HealthBethesdaUSA
  2. 2.Laboratory Animal Sciences Section, National Institute of Diabetes, Digestive and Kidney DiseasesNational Institutes of HealthBethesdaUSA
  3. 3.Flow Cytometry Core, National Heart, Lung and Blood InstituteNational Institutes of HealthBethesdaUSA

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