The pGLO Kit SDS-PAGE Extension is designed to be performed following the pGLO Bacterial Transformation Kit. The extension is designed to identify and visualize green fluorescent protein (GFP) from the proteome of transformed E. coli, which also contains thousands of other proteins. Because only the cloned GFP glows, students can use this phenotypic trait to quickly identify the GFP within the complex mix of proteins.



In the first part of the exercise, students use the pGLO Bacterial Transformation Kit to transfer pGLO plasmid encoding GFP into E. coli, a common prokaryotic organism used for DNA propagation and protein expression. Colonies of E. coli are qualitatively examined for fluorescence to determine whether the pGLO gene is being expressed. In the second part of the laboratory activity, sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) is employed to separate the entire repertoire of proteins expressed in E. coli, including the foreign GFP responsible for transferring the fluorescence trait.