Ethanol Fixation and DAPI Staining: A Method to Visualize DNA in C. elegans
Published: April 30, 2023
Abstract
Source: Clarke, E. K., et al. Manipulation of Ploidy in Caenorhabditis elegans. J. Vis. Exp. (2018).
Here, we describe a technique used to visualize genomic DNA in fixed, intact nematodes. The video includes an example protocol to count chromatin in unfertilized C. elegans oocytes.
Protocol
This protocol is an excerpt from Clarke et al, Manipulation of Ploidy in Caenorhabditis elegans, J. Vis. Exp. (2018). Tetraploid strains can be validated by counting the number of chromosomes in their unfertilized oocytes. Fluorescent microscopy can be used to screen for the number of chromosome pairs in unfertilized diploid oocytes (prior to the meiotic division…
Materials
| Dissecting Microscope | Motic Microscopy | SMZ171 | |
| Name | Company | Catalog Number | Comments |
| Staining | |||
| Hoechst 33258, Pentahydrate | Biotium | 40045 | |
| DAPI | Biotium | 40011 | |
| Name | Company | Catalog Number | Comments |
| Ethanol Fixation | |||
| Ethanol, Pure, 190 Proof (95%), USP | Koptec, VWR | 89125-166 | |
| Name | Company | Catalog Number | Comments |
| M9 Buffer | |||
| Sodium chloride, Biotechnology Grade | VWR | 97061-278 | |
| Potassium phosphate, Monobasic Anhydrous Grade | VWR | 97062-346 | |
| Sodium phosphate, monobasic dihydrate | Fisher Scientific | AC271750025 |
Citer Cet Article
Ethanol Fixation and DAPI Staining: A Method to Visualize DNA in C. elegans. J. Vis. Exp. (Pending Publication), e20136, doi: (2023).