I am isolating plasmid DNA manually using alkaline lysis method. Everytime after precipitating with isopropanol and ethonal, along with DNA pellet some other white colour precipitates observed. Also the concentration of the plasmid on nano-drop spectrometer comes more than 2000-3000ng/ul but when run on a gel the concentration seems to be very less even the bands are on clearly visible.
Can anyone suggest me solution for this.
You can try the cultures which is pre-amplified with Chloramphenicol or any suitable antibiotics. Possibly, the spectral data can be varied according to the structural confirmation of plasmid.
Hi Madhvi!
You can check for RNA contamination. Usually in manual preps if the RNase treatment is not proper you will get RNA contamination in the precipitated DNA. This gives you increased nanodrop readings as well. The contaminating RNA would be visible in agarose gel when you run DNA. If you do not run out the dye, it will appear as diffused band way below the original plasmid band.
Hope this helps.
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