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    Protocol collection: Dissociation and FACS isolation of embryonic and post-embryonic C. elegans intestine cells for RNA-seq analysis

    Robert TP Williams, Erin Osborne Nishimura

    Published: 2022-09-26 DOI: 10.17504/protocols.io.5jyl895jdv2w/v1
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    Abstract

    This is a collection of protocols for the isolation of C. elegans intestine cells through FACS from embryo, L1, and L3 stage . The protocols are focused on collecting material for RNA-seq analysis, but could be utilized for other biochemical assays. Troubleshooting suggestions are listed at key steps of the protocols.

    To perform this experiment, perform the protocols in the following order:

    1. Synchronized C. elegans culture on NGM plates for FACS isolation of intestine cells

    2. [Embryo, L1, or L3] stage C. elegans dissociation for FACS isolation and RNA-seq analysis of intestine-specific cells

    3. FACS isolation of intestine-specific C. elegans cells

    A single experiment cycle takes approximately two weeks to complete.

    Steps

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