How to isolate Plasmid DNA
Автор: Genomics Lab
Загружено: 2013-09-27
Просмотров: 26808
Описание: In this video, Plasmid DNA is being extracted from the Bacterial culture. Bacteria containing plasmid are harvested from liquid bacterial culture by centrifugation. The suspension of Bacteria is made in isotonic solution containing a detergent, Sodium dodecyl Sulphate(SDS), and an alkali namely Sodium Hydroxide(NaOH). SDS lyses the cells and denatures the proteins, alkali condition aids in denaturing the genomic DNA and plasmid DNA. Lysed cells are further neutralised by Potassium acetate (pH 5.2), this helps to renature the plasmid DNA and Genomic DNA. Precipitate is separated by high speed centrifugation. Plasmid from the supernatant is recovered by precipitation using isopropanol or ethanol.The presence of Plasmid DNA is confirmed by performing Gel Electrophoresis. The formation of band proves the presence of Plasmid DNA in the Bacterial culture.
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