Hi everyone
I isolated some plasmids, using alkaline lysis method (which I have been following for 2-3 years now) and the final plasmid pellet after washing in 70% ethanol was dried. Then I added 50 ul of miliQ water to each pellet and stored the vials at -20 degrees overnight. Next day when i thawed the plasmid vials to quantify the DNA, I noticed that the pellets were not dissolved, instead a white precipitate was there- I took nanodrop reading and the concentration was in the range from 3300-4200 ug/mL and 260/280 around 1.8-1.88; 260/230 around 2-2.28.
Suggestions will be helpful as I have to go for transfection with these plasmids.
Thanking you in advance
It sounds like your plasmid did dissolve just fine in water, otherwise you would not have such a high concentration reading with the nano drop. The precipitate is probably some salt or other molecule that is less soluble. You could try and and just pellet the ppt to remove it and double check the solution again.
I suggest you increase your final vol maybe your con of DNA is more than this.
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