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A simple and effective method to isolate germ nuclei from C. elegans for genomic assays

View ORCID ProfileMei Han, Guifeng Wei, Catherine E. McManus, View ORCID ProfileValerie Reinke
doi: https://doi.org/10.1101/371351
Mei Han
1Department of Genetics, Yale University, New Haven CT 06520, USA
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Guifeng Wei
2Developmental Epigenetics, Department of Biochemistry, University of Oxford, Oxford OX1 3QU, UK
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Catherine E. McManus
1Department of Genetics, Yale University, New Haven CT 06520, USA
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Valerie Reinke
1Department of Genetics, Yale University, New Haven CT 06520, USA
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Abstract

The wide variety of specialized permissive and repressive mechanisms by which germ cells regulate developmental gene expression are not well understood genome-wide. Isolation of germ cells with high integrity and purity from living animals is necessary to address these open questions, but no straightforward methods are currently available. Here we present an experimental paradigm that permits the isolation of the nuclei from C. elegans germ cells at quantities sufficient for genomic analyses. We demonstrate that these nuclei represent a very pure population and are suitable for both chromatin immunoprecipitation (ChIP-seq) and transcriptome (RNA-seq) analyses. The method does not require specialized transgenic strains or growth conditions and can be readily adopted by other researchers with minimal troubleshooting. This new capacity removes a major barrier in the field to dissect gene expression mechanisms in the germ line of C. elegans. Consequent discoveries using this technology will be relevant to conserved regulatory mechanisms across species.

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The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Posted July 17, 2018.
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A simple and effective method to isolate germ nuclei from C. elegans for genomic assays
Mei Han, Guifeng Wei, Catherine E. McManus, Valerie Reinke
bioRxiv 371351; doi: https://doi.org/10.1101/371351
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A simple and effective method to isolate germ nuclei from C. elegans for genomic assays
Mei Han, Guifeng Wei, Catherine E. McManus, Valerie Reinke
bioRxiv 371351; doi: https://doi.org/10.1101/371351

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