Biopharma Insights This method allows for the generation of tetraploid and triploid Caenorhabditis nematodes from any diploid strain. Polyploid strains generated by this method have been used to study chromosome interactions in meiotic prophase, and this method is useful for examining important basic questions in cell, developmental, evolutionary, and cancer biology.
Total time
~16 days (including RNAi induction, strain isolation, and verification)
Model organism
Caenorhabditis elegans
Steps
1
Set up rec-8 RNAi feeding protocol
Prepare RNAi bacterial strains and culture conditions over Days 1–3 to initiate rec-8 knockdown in target C. elegans populations.
▶ 01:25
2
Generate and isolate tetraploid strains
Monitor nematode populations Days 4–16 for polyploid formation following rec-8 RNAi induction, then select and isolate tetraploid individuals.
▶ 02:32
3
Verify ploidy status with DAPI staining
Stain isolated strains with DAPI and use fluorescence microscopy to confirm tetraploid nuclear DNA content relative to diploid controls.
▶ 03:57
4
Confirm successful tetraploid strain establishment
Present representative DAPI-stained images and fluorescence data demonstrating increased nuclear ploidy in generated strains.
▶ 05:56