Plasmid DNA extraction
Dear colleagues, I am trying to extract plasmid DNA from salmonella enterica isolates using alkaline lysis method, but I have a big problem with the resuspension of plasmid precipitate after, isopropanol precipitation, as the precipitate is very difficult to be dissolved with TE buffer without pipetting or vigrous shaking that lead to plasmid damage.
I am very appreciate if there is anyone can help
Dissolve in TE buffer at 37 degrees with shaking overnight in an incubator.
Also, ensure that you have allowed time for the isopropanol to dry off before adding TE. Sometimes washing the precipitate in Ethanol after the isopropanol step can exchange the Isopropanol for more volatile Ethanol which will dry off quicker.
If you are using PEG fractionation later on then a little DNA shearing from a poke with a pippette should be easy enough to remove further downstream.