I have a dissolved plasmid pellet in water. Can I pellet it again by centrifuging it at 13000 rpm? If not, why?
3
-
$\begingroup$ Only by centrifuging? $\endgroup$– Chris ♦Commented Mar 9, 2020 at 7:43
-
$\begingroup$ Is this a homework question? have you done any research on this? Could you share your findings here? $\endgroup$– AliceD ♦Commented Mar 9, 2020 at 8:03
-
$\begingroup$ Hi Alex, It wasn't homework question. After centrifuging a while, the concentration of the plasmid reduced a bit. (1.7 ug to 1.35 ug). Couldn't find a proper literature on it. So I thought it is worth asking. $\endgroup$– RKKCommented Mar 11, 2020 at 6:03
Add a comment
|
1 Answer
$\begingroup$
$\endgroup$
0
No, you cannot pellet dissolved DNA with ultracentrifugation.
Yes, you can recover a pellet with additional treatments, similarly to how you got it in the first place; only instead of your input being homogenized cells, or tissue, or extract, or whatever you used to obtain your DNA, it would now just be your aqueous solution of DNA.
For instance, you may have performed a TRIzol extraction to separate your DNA into an aqueous phase, prior to spinning it through mini prep columns to elute your purified DNA in water. You could perform the procedure again and you'd have your pellet once more after the final alcohol (ethanol or isopropanol) precipitation step.
The key idea with pelleting DNA (or any nucleic acid) is to make it insoluble; this is achieved with by using alcohols and salts. If it is soluble, nucleic acids cannot 'come out of solution' to form a pellet.
