Hello everyone,
My question for sure is silly, but I have been arguing with my labmate about this. If our plasmid concentration is 0.776 µg/µL and he wants to make a final concentration for transfection of 1 µg/µL and 6 µg/µL, how is it possible to achieve a concentration of 6 µg/µL from 0.776 µg/µL?
Using the equation C_1 V_1 = C_2 V_2 :
v2 = 50 ul
c2= 6 ug/ul
Please help me understand. I have a brief knowledge of chemistry and calculation
You can't make a final concentration that is higher than your initial concentrations. If you really need a higher concentration then you would need to concentrate your DNA by precipitation and resuspending in a smaller volume.
However I am not sure you need the higher concentration. Do you perhaps mean that you need 1µg or 6µg of plasmid for the transfection (but not per µl)?
Hi!
I'm not sure that is possible. For me, you can't diluted an initial solution to have something more concentrated. I suggest you to concentrated your initial plasmid solution, by lyophylizing it a little bit, an measured the concentration to have something more concentrated. If it success, you can now apply your formula recommendated for dilution.
Ryan .. Hi!, First of all, no questions are silly, every question is a learning point.
Next, I would agree with Michael J. Benedik that you cannot get a higher concentration by diluting already obtained plasmids at a lower concentration. You can try precipitating it again and resuspend it in lower volume.
Ryan, you can use 6 ug by using a larger volume, you would need about 8 ul of your plasmid prep.
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