Hi, I use DMEM that was been as complete growth medium (CGM). If working volume for 24 well plate is 1ml, how can I distinguish between how much volume of CGM (grwoth medium) and treatment sample solution should be added to well plate? If I put the different volume for different concentration of treatment solution, absolutely the CGM volume will differ (based on M1V1=M2V2 calculation). Is it fine to have low volume of CGM (growth medium), to the cell growth?
Hi Samiihah,
I hope I understand your question: When you add substances to one well to study its effect, you have to compare it with a well that received the same amount of solvent (water, buffer, DMSO ,...). In addition, an untreated well will show you whether the solvent has an effect by its own.
Hi Uwe,
You explanation is right. My sample is in liquid. Then, I need to put the sample liquid into well together with DMEM. Different concentration of sample liquid will have the different volume of its, to be put in well. Does the volume of DMEM effected, Can I have different volume of DMEM (growth medium); but having same working volume for each well?
I would just add 1 ml DMEM to each well. When you add, e.g. 10µl to 100 µl of your substance to individual wells prepare wells with the same volume of solvent. As long as the cells tolerate the solvent you are fine. You already have your control for the dilution effect.
However, you want to keep the volume as small as possible. If you need to add large volumes to see an effect, you should start to concentrate your substance ( e. g. freeze/dry).
As soon as you know that your solvent has no big effect you may treat all wells with the same volume but different concentrations of your substance.
Does the concentration of sample will be diluted after added to the DMEM?
Your control samples should have an equal volume of diluent of the same solvent as your experimental treatment.
Hi, I agree with Mr.Uwe. Moreover, I would like to tell you that if your treatment in a well is of 100ug/ml concentration, you can take 10ul from 1000ug stock and add to the well containing cells in 990ul of DMEM. I hope you got clarified.
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