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Escherichia Coli Transformation Lab Report

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Transformation Escherichia coli was first described by Mandel and Higa ( Yasuhiko et. al, n.d.) , when E.coli were treated with calcium chloride, it will neutralize the bacterial cell to make it competent. The prolonged exposure of cells to CaCl2, substitution of Ca2+ ions by other cations, such as Rb+, Mn2+, and K+ can improve the efficiency of transformation. Purification requires the removal of protein and isolation of the DNA by calcium chloride gradient. Alkaline lysis method is used for plasmid DNA extraction. Then, lyse the bacteria with a solution containing sodium dodecyl sulfate (SDS) and sodium hydroxide and the chromosomal DNA and plasmid DNA will be denatured. The chromosomal DNA and proteins precipitate in a complex and then

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