Sep 20th (Prof. Stephane's lab)
- Justin: If lysis method works with gram positive, its likely to work with fungal cells
AudioLyse - should work on gram positive bacteria for fungal cells to work
Atomization should also work on fungi as well since its gram positive bacteria
FTA paper with eDNA —> hows that gonna work?
- Justin: ITS region is a widely studied barcode gene for fungi, good choice
- bypass amplification step, only CRISPR reaction; identify the detection limit
- Run a negative control on a piece of fungal DNA; make sure it doesn't amplify fungi and cross-species
- run the assay with yeast; maybe reach out to professor kourosh and alexandra (his postdoc) - their main research is algae
- test with soil; it might contain chytrid
For quantitative results: use a standard curve?
- extraction step efficiency need to be consistent
SUBMERGE YOUR SWAB IN LIQUID THATS ALL YOU NEED. NOT TOO MUCH CUZ THAT WILL DILUTE
- concentration by evaporation - would require centrifuge
can check with swab from professor Stephane's lab - whether they have chytrid in it via PCR assay
- time is more important than cost
Ideally, should be scaled up to 30ish samples
- 1-2 batches test
- minimize the handling would be better
- sample collection at night are standard; frogs are nocturnal