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Large-scale sorting of C. elegans embryos reveals the dynamics of small RNA expression

MPG-Autoren
http://pubman.mpdl.mpg.de/cone/persons/resource/persons73941

Li,  Na
Computational Epigenetics (Ho-Ryun Chung), Independent Junior Research Groups (OWL), Max Planck Institute for Molecular Genetics, Max Planck Society;

http://pubman.mpdl.mpg.de/cone/persons/resource/persons50123

Chen,  Wei
Dept. of Human Molecular Genetics (Head: Hans-Hilger Ropers), Max Planck Institute for Molecular Genetics, Max Planck Society;

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Zitation

Stoeckius, M., Maaskola, J., Colombo, T., Rahn, H.-P., Friedländer, M. R., Li, N., et al. (2009). Large-scale sorting of C. elegans embryos reveals the dynamics of small RNA expression. Nature Methods, 6, 745-751. doi:10.1038/nmeth.1370.


Zitierlink: http://hdl.handle.net/11858/00-001M-0000-0010-7D13-D
Zusammenfassung
Caenorhabditis elegans is one of the most prominent model systems for embryogenesis, but collecting many precisely staged embryos has been impractical. Thus, early C. elegans embryogenesis has not been amenable to most high-throughput genomics or biochemistry assays. To overcome this problem, we devised a method to collect staged C. elegans embryos by fluorescence-activated cell sorting (eFACS). In a proof-of-principle experiment, we found that a single eFACS run routinely yielded tens of thousands of almost perfectly staged 1-cell stage embryos. As the earliest embryonic events are driven by posttranscriptional regulation, we combined eFACS with second-generation sequencing to profile the embryonic expression of small, noncoding RNAs. We discovered complex and orchestrated changes in the expression between and within almost all classes of small RNAs, including microRNAs and 26G-RNAs, during embryogenesis.