Chromosome Spreads with Centromere Staining in Mouse Oocytes
This chapter describes a technique for performing chromosome spreads from mouse oocytes. It is based on a previously described protocol (Hodges and Hunt, Chromosoma 111: 165–169, 2002), which we have modified. Chromosomes are stained with either Propidium Iodide or Hoechst. This spreading technique allows for simultaneous immunostaining of proteins associated with chromosomes. It is very useful to stain spreads with CREST serum which labels kinetochores, to be able to distinguish bivalents (chromosome pairs), dyads or univalents (paired sister chromatids), and single sister chromatids without ambiguity.
Key wordsChromosome Spreads CREST Meiosis Bivalent Univalent Sister chromatid Cohesion Mouse oocytes
We thank S. Touati for comments on the manuscript. K.H. was supported through a PhD fellowship by the Ministère de la Recherche and ARC. Research in the group of K.W. is supported by a grant from La Ligue Nationale Contre Le Cancer (Comité d’Ile de France), the UPMC and CNRS.
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