I need help the the steps and the math required to dilute a concentration of cells (4.0x10^5 cells/mL) across 6 wells.
I would like to have increments of .5x10^5 (50,000) so the first well will have 4.0x10^5, the second well will have 3.5x10^5, the third with 3.0, and so on. Each well should have 2.0 mLs of the cell suspension, so i'll likely have around 12ml + ~5-10mL of buffer in order to perform the dilution.
Can someone explain a simply way of doing this? Basically I will attain the first concentration by doing a calculation and taking an automated cell count, but I want a quick yet assurable way to attain lower increments of cells/ml without having to perform six different counts.
You should make a stock solution of your cells of the highest conc i.e, 4.0x10^5 cells/ml. To find out how much ml of the stock you need and how to obtain each concentration do the following calculations for each concentration:
[stock] / [required] = Dilution factor(DF)
Vol required from stock = total volume / DF
For example, in your case
for the first concentration you add 2ml to the well from the stock (4.0x10^5cells/ml)
for the second conc i.e, 3.5x10^5 cells/ml
D.F = 4.0 / 3.5 = 1.142857
Volume req from stock (in uL) = 2000 uL / 1.142857 = 1750 uL
Therefore you take 1750 uL from the stock and make it up to 2ml with buffer
Do the same calculations for each concentration and you will get the req volume you need to take from the stock for the different conc in the order 2000, 1750, 1500, 1250, 1000 & 750 uL respectively. Make them up to 2ml with the buffer. Add them up before (2000, 1750, 1500, 1250, 1000 & 750 uL) making the stock to find out how much of the stock you need to make.
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