Question

in the plasmid purification procedure, what does the neutralization soulation achieve?

Answer 1

Ans:- in plasmid isolation strong alkaline solution od sodium dodecyl sulfate and NAOH is added to cell mixture. this alkaline solution will disrupt cell membrane and DNA including chromosome DNA and plasmid DNA. this condition will increase Ph of cell mixture. strong Neutralizing buffer like potassium citrate added to neutralize high alkaline condition in cell mixture. this Neutralizing action will facilitate renaturing the plasmid DNA. additionally potassium citrate precipitate sodium dodecyl sulfate hence remove detergent from cell mixture. cell mixture is then centrifuge this time pellet contains cell debris which was lysed by alkaline solution and supernatant liquid with plasmid is isolated and taken to gel electrophoresis.