Document Detail

Methods for meiotic chromosome preparation, immunofluorescence, and fluorescence in situ hybridization in Daphnia pulex.
MedLine Citation:
PMID:  19685328     Owner:  NLM     Status:  MEDLINE    
The genus Daphnia has an intriguing reproductive mode of cyclical parthenogenesis. This reproductive mode has been studied for centuries, but cytogenetic information is lacking due to technical limitations of classical methods. We have developed methods for the preparation and examination of meiotic chromosomes of Daphnia pulex from oocytes and spermatocytes. Oocyte chromosome preparations are obtained by isolating individual oocytes after the release of yolk granules from the ovary using pressure and capillary action. Spermatocyte chromosomes are prepared using a conventional squash method. Cryosectioning is an easy and fast way to prepare sections. We also illustrate the application of immunofluorescence staining against alpha tubulin, as well as fluorescence in situ hybridization (FISH) using the intergenic spacer of ribosomal DNA or single-copy cosmid clones.
Dai Tsuchiya; Brian D Eads; Miriam E Zolan
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Publication Detail:
Type:  Journal Article; Research Support, U.S. Gov't, Non-P.H.S.; Review    
Journal Detail:
Title:  Methods in molecular biology (Clifton, N.J.)     Volume:  558     ISSN:  1064-3745     ISO Abbreviation:  Methods Mol. Biol.     Publication Date:  2009  
Date Detail:
Created Date:  2009-08-17     Completed Date:  2009-10-21     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9214969     Medline TA:  Methods Mol Biol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  235-49     Citation Subset:  IM    
Department of Biology, Indiana University, Bloomington, IN, USA.
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MeSH Terms
Chromosomes / chemistry,  ultrastructure
Cytological Techniques / methods*
Daphnia / cytology*,  genetics*
Fluorescent Antibody Technique / methods
In Situ Hybridization, Fluorescence / methods*
Meiosis / genetics,  physiology*

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