Jul 25, 2019
Version 2
Spot-bleaching of a handful of C. elegans nematode worms V.2
- 1University of Cambridge
Protocol Citation: Cristian Riccio 2019. Spot-bleaching of a handful of C. elegans nematode worms. protocols.io https://dx.doi.org/10.17504/protocols.io.5ueg6te
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: July 25, 2019
Last Modified: July 25, 2019
Protocol Integer ID: 26214
Abstract
This protocol describes the spot-bleaching of C. elegans
Guidelines
To get rid of contaminants, you can bleach your worms in a tube (most efficient method) or spot bleach on an agar plate (faster method).
Materials
MATERIALS
M9 solution for nematode culture
bleaching solution for C. elegans
Handful of gravid (pregnant) C. elegans worms
Leica L2 binocular microscope
Safety warnings
Wear a lab coat and gloves when you handle the bleaching solution. Avoid wearing gloves next to a flame.
Before start
Prepare bleaching solution. Make sure the bleaching solution is less than one month old.
1. Put a drop (20 to 50 µl) of bleaching solution on the edge of a clean NGM plate seeded with E. coli .
2. Pick several gravid hermaphrodites in the drop. The bleaching solution will kill the contaminants and hermaphrodites but will soak into the plate before the embryos hatch.
3. The next day the L1 larvae will have crawled into the E. coli OP50 lawn. Transfer them to a clean NGM plate seeded with an E. coli OP50 lawn, or cut the patch of dead bacteria/bleach agar.