May 17, 2023
Adding 1-hydroxyphenazine to L4 C. elegans in liquid culture
- Muhammad Zaka Asif1,
- Man Shah1,
- Yosef Smadi1
- 1Edison Lab, UGA
Protocol Citation: Muhammad Zaka Asif, Man Shah, Yosef Smadi 2023. Adding 1-hydroxyphenazine to L4 C. elegans in liquid culture. protocols.io https://dx.doi.org/10.17504/protocols.io.yxmvm28png3p/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: May 17, 2023
Last Modified: May 17, 2023
Protocol Integer ID: 82036
Abstract
This protocol describes day 4 of our workflow to grow worms in liquid culture to induce the production of natural products (in this case, 1-HP derivatives). In this protocol, we add the toxin to our worms grown in liquid culture to induce the production of glycosylated products.
Follow steps 1-6 from the protocol for transferring C. elegans to S-buffer, but centrifuge at 525 RCF for 1 minute
Dilute to a final concentration of 30,000 worms/mL in S-basal and final toxin concentration of 22.3 µM 1-HP.
Repeat steps 1-2 for 1.1% DMSO and bacteria-only controls.
If using negative controls, repeat steps 1-3 for them.
Put the flasks back in the shaker and allow to incubate for 24 hours.