Adding Molecular Biology to the Biochemistry Laboratory

Villanova University, Villanova PA

Investigators

Abstract

Chemistry (12) Experiments in molecular biology are added to the students' biochemistry laboratory experience to reflect state-of-the-art techniques and procedures. In addition to protein isolation and characterization, enzyme kinetics, and carbohydrate or lipid isolation and characterization, emphasis is placed on such techniques of modern molecular biology as DNA isolation, gene isolation and cloning, the polymerase chain reaction, cell transfection and gene expression. The following molecular biology experiments are among those being adapted from standard literature and implemented in the Biochemistry laboratory. Bacteria are transformed with a plasmid including the gene for green fluorescent protein (GFP). The GFP is isolated using procedures from standard literature and characterized by electrophoresis and fluorescence spectroscopy. The identity of GFP is confirmed using an immunoassay with anti-GFP polyclonal antibodies. The plasmid with the GFP gene is isolated from the bacterial culture and characterized by restriction analysis. Some of the isolated plasmid is reserved to transform cells at the beginning of the semester that follows. An ultracentrifuge is used for DNA and plasmid isolation. A plate reader is used to read the results of the immunoassay and for enzyme kinetics experiments. Multipipettors are used to facilitate the completion of the plate assays.