Dissolving DNA in TE buffer - troubleshooting - (May/06/2005 )
I have extracted genomic DNA from whole blood sample using an ethanol precipitation method and then dissolved the pellet in TE buffer.
After a week that I was off, the DNA seems still not to have been fully dissolved in all samples.
I’ve just realized today that I was using from the beginning a 10x TE buffer to dissolve DNA instead of 1x TE buffer and I’m wondering now whether the excess of salt amount is a factor that prevents DNA to dissolve, or the cloud I see is salt and not DNA?
Thanks for any reply!
When you warm up your samples, does the cloud diappear? If yes, I think that is the salt.
As I didn't know what to do, I decided to run the precipitation step again and dissolve the DNA pellet in a TE buffer solution with the correct concentration this time. After overnight incubation in 4oC, I was happy to find out that I didn't see clouds anymore. Also the measurement with spectrophotometry gave good results.
So I don't have the opportunity to check the results after warming up the samples, but I will certainly keep this in mind for any further similar problem that I might get.
Thanks a lot for your answer!!
sounds like you were able to rectify the problem yourself, the chances were that what you saw was excess salt and the extra precipitation step removed this. Doesn't hurt to go the extra mile to get clean DNA