Adding 1-hydroxyphenazine to L4 C. elegans in liquid culture
Muhammad Zaka Asif, Man Shah, Yosef Smadi
Published: 2023-05-18 DOI: 10.17504/protocols.io.yxmvm28png3p/v1
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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.
Steps
1.
Follow steps 1-6 from the protocol for transferring C. elegans to S-buffer, but centrifuge at 525 RCF for 1 minute
2.
Dilute to a final concentration of 30,000 worms/mL in S-basal and final toxin concentration of 22.3 µM 1-HP.
3.
Repeat steps 1-2 for 1.1% DMSO and bacteria-only controls.
4.
If using negative controls, repeat steps 1-3 for them.
5.
Put the flasks back in the shaker and allow to incubate for 24 hours.
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