I have to do a clonogenic and a mtt assay to test the toxicity of two different drugs accross three cell lines. Im struggling to get to grips with determining the cell numbers to plate out initially before any drug is added to the plates.as well as how to lay these out (kind of left to get on with it and it is my first time doing both assays ) different protocols online are really confusing me so I am sorry in advance for any noob questions of anything I may have overlooked.
So, after i have trypsinized my cells and it is time to resuspend my pellet, how much media do i resuspend in? or does this not matter? and once I have my cell count, do I treat my re-suspended pellet suspension as my cell stock, and take however many cells I need from there?
Lets say I want to put a 1000 cells in each well (of a 96 well plate), to work out how much I need to take from my stock do i divide the number of cells I want over the number of cells I have counted and mulitply this by the volume I resuspended in followed my x by 1000 to get it into ul?
Any help or advice will be greatly appreciated