Collection Citation: Robert TP Williams, Erin Osborne Nishimura 2022. Protocol collection: Dissociation and FACS isolation of embryonic and post-embryonic C. elegans intestine cells for RNA-seq analysis. protocols.io https://dx.doi.org/10.17504/protocols.io.5jyl895jdv2w/v1
License: This is an open access collection distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this collection and it's working
Created: January 19, 2022
Last Modified: September 26, 2022
Collection Integer ID: 57152
Keywords: C. elegans, intestine, FACS, single cell suspension, cell dissociation
Funders Acknowledgement:
National Institutes of Health
Grant ID: R35GM124877
Bridge to Doctorate at Colorado State University
Grant ID: 1612513
National Science Foundation
Grant ID: 2143849
Webb-Waring Biomedical Research Award
Grant ID: E.O.N.
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.
