Isolation, Cloning, and Translation of Plasmid DNA
Abstract:
The http://professional-academic-writing-services832.mystrikingly.com objective of this experiment was to clone a kanamycin gene into the MCS of a pUC18 plasmid, and then to transform cells with the plasmids. Purified pUC18 and pKan plasmid samples were obtained. A 0.7 % agarose gel was prepared, and the wells loaded with the plasmid samples. Restriction endonucleases were used to cut a kanamycin resistance gene from a pKan plasmid. DNA ligases were used to ligate the kanamycin resistance gene on to the multiple cloning site of the pUC18 plasmid. Escherichia coli (strain DH5α) were then transformed with plasmids. The presence of the kanamycin resistance gene in the pUC18 was determined using the indirect (pUC18 selection) and direct selection methods. The results from the gel image were inadequate. Zero colony counts were recorded on the kanamycin plates for the indirect selection method. Zero colony counts were recorded on the kanamycin/ carbenicillin plate for the direct selection method.
