Hi everyone. I would like to know a good protocol for RNA extraction and real time PCR in C. elegans. In particular, I am asking how many worms are needed for efficient extraction?
Check these protocoles :
Article High-Throughput Quantitative RT-PCR in Single and Bulk C. el...
https://www.zamanianlab.org/ZamanianLabDocs/protocols/Caenorhabditis_elegans/RNA_Extraction/RNA_Extraction/
https://greerlab.wustl.edu/items/c-elegans-rna-extraction/
You can use exactly 1 worm, but most folks want to start with a "pellet" of worms that's about 100 microliters.
In the past, we used 1 plate of adults after doing an egglay for 2 hours for synchronization. It gives you about 200 worms or so. For starting I would try 2-3 plates of adult animals if that is what you are interested in.
- qPCR of a handful of genes: ≥100–500 worms (L4/young adults).
- RNA-seq (standard bulk kits needing 50–200 ng total RNA input): ~1,000–5,000 worms.
- Low-input RNA-seq kits (5–10 ng input): ~300–1,000 worms.
- Single-worm or