Mar 31, 2026

Public workspaceSychronisation of C. elegans

DOI
https://dx.doi.org/10.17504/protocols.io.14egn514yg5d/v1
  • Ella Fiebig1
  • 1The University of New South Wales, Kensington, NSW, Australia
Ella Fiebig
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DOI: https://dx.doi.org/10.17504/protocols.io.14egn514yg5d/v1
Protocol Citation: Ella Fiebig 2026. Sychronisation of C. elegans. protocols.io https://dx.doi.org/10.17504/protocols.io.14egn514yg5d/v1
License: This is an open access protocol 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 protocol and it's working
Created: March 31, 2026
Last Modified: March 31, 2026
Protocol Integer ID: 314151
Keywords: using stiernagle, sychronisation, protocol
Abstract
Protocol designed using Stiernagle (2006) with modifications.
Troubleshooting
Washing
Wash C. elegans from 3-5 plates using 3mL of M9. Ensure plates contain a mixed population of C. elegans with gravid nematode adults.
Wash C. elegans by centrifuging 800 x g for 2 minutes. Remove supernatant and resuspend in M9. The worms should form a pellet while the bacteria remain suspended in the supernatant.
Repeat washing step 3x to remove bacteria from C. elegans.
Bleaching
Add 4mL of freshly made bleaching solution
Bleach solution:
NaCIO: 3mL
5M NaOH: 1mL
H2O: 1mL
Shake constantly for 3-5 minutes but not longer than 5 minutes. This causes the eggs to be released from the gravid c. elegans.
Stop the reaction by adding 40 mL of M9 buffer.
Quickly centrifuge at 1300xg for 3 minutes and discard supernatant.
Repeat wash step 2 more times using an equal amount of M9 buffer.
After the final wash, transfer the eggs onto the NGM plates and incubated at 25 C°. The eggs should develop to the L2 phase after 24 hours, then they can be supplemented with OP50 E. coli to promote growth to the L4 phase.
Protocol references
Stiernagle, T. (2006). Maintenance of C. elegans. In C. elegans Research Community (Ed.), WormBook: The Online Review of C. elegans Biology, WormBook. Available at http://www.wormbook.org/chapters/www_strainmaintain/strainmaintain.html.