I've set up an experiment with C2C12 cells where I have transfected them with Smn siRNA for 48 hours to knock down Smn expression. I want to observe the effects of prednisolone on these cells at 700nM concentration. However, the issue I am facing is that I cannot change the siRNA-media and replace it with a 700nM prednisolone-media solution from C1V1=C2V2. Instead, I have to directly add my 7 microM prednisolone stock solution to the siRNA media. The problem I have encountered is how do I calculate the volume of 7 microM prednisolone to add to 2ml of media to get a concentration of 700nM?
Dear Joseph,
You Have = [stock of 7uM (micromolar)]
You Want = [final] of 700nM
The Final volume of 2mL of media
Thus:
( 700nM x 2mL ) / 7uM = x
x = 200uL
Add 200uL of 7uM stock to 2mL of medium to have a final concentration of 700nM. Mix well.
Best Wishes!
I have same answer.
7000nM X x = 700nM X 2000 ul
x= 1400000/7000
=200ul. Therefore you add 200uL from your 7uM (7000nM) stock to 2ml medium.
You can confirm the same from Molarity calculator or any other calculator available online.
Good luck !!!
Sorry, but why are you not able to remove siRNA-containing medium?
SiRNA needs some hours to days to led to reduced protein expression (maybe leading to significantly reduced activity of your protein of interest). Moreover, in my opinion, you should never ever leave cell for more than 24 h in siRNA-containing medium when using lipofection.
Thus, could you give us more information regarding your experimental style. After that it would be more useful to answer your question.
In any case, your calculation seems ok.
While u add the transfection media keep it to 1800 final and add 200 ul of ur drug directly. This dilution is not good for cells. I would suggest making higher concentrations like 7milimolar. So u would only need 2 ul for 2ml.
For future, you can also use this Dilution Calculator, which is really easy to use! https://www.hellobio.com/dilutioncalculator/
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