I want to test the toxic effects of a plant alkaloid on yeast. Apart from checking OD (growth rate) after different time intervals treating with the alkaloid, what other kinds of experiments can I do? Can I do trypan blue assay for yeasts?
Dear Rupam,
Yes you can do tryphan blue staining for checking the viability. Also, you can do methylene blue staining too. Apart from this staining, the designing of other experiments depends on your project objectives and other facilities that you have currently.
Do some literature and look at other similar kind of papers to get an idea about other experiments.
Good Luck..!!!!
Dear Upendar Sir,
The aim is to find the toxic effects on yeast cells and compare them with mutants.
Dear Rupam
Why don't you try MTT assay'. Its much mare sensitive and will give you reading of viable cells only.
Tried both MTT and XTT. MTT is more simple and cheaper than XTT. But XTT is better, I agree that
Colony forming unit assay is the easiest way. Treat a yeast culture of known cell density with a growth inhibitory concentration of your compound, remove an aliquot (approximately 100ul) at different times (I usually do 30 min, 1 h, 2 h, 4 h, 6 h) and spread it on a YPD agar plate. Incubate for 2 days at 30 degrees then count colonies that have formed. Because you know the cell density you can calculate how many colonies should form if there is no cytotoxicity, anything less than this number in the treated samples suggests there is cytotoxicity. Tetrazolium based assays are not that appropriate in yeast since they rely on respiration, this will be low in glucose containing media, also false positive results are possible due to an inhibitory effect of your compound on respiration. We did something similar with the compound TA-289, have a look on my profile for it.
- Badges
- Science topic
- Similar topics
- Biological Science
- Molecular Biology