Recombinant DNA techniques played a central role in the recent emergence of biology into the golden age. This program enables the advanced student to approach these frontiers in biotechnology. The program provides a challenging series of seven 3-hour laboratory sessions which are intended to give students hands-on-experience and a detailed understanding of these new investigative techniques and their potentials.
In the program, students prepare a library of recombinant plasmids which contain different regions of the lambda phage genome. They then amplify and characterize the lambda DNA in selected plasmids in order to identify the phage genes that they contain. The basic strategy followed in the program is shown on the facing page and is one that has been used by investigators around the world to clone DNA molecules from literally thousands of prokaryotic and eukaryotic organisms. However, many of the procedures have been modified to make them safe and suitable for a college laboratory that meets 3 hours once per week. The program provides essentially all of the instructions, chemicals, sterile media, and expendable accessories that are needed to carry out DNA electrophoresis, restriction nuclease digestions, DNA ligations, bacterial transformations, bacterial selection for ampicillin resistance and ß-galactosidase production, plasmid isolation, DNA hybridization, and Southern blot procedures. A centrifuge that can be operated at a force of at least 3,000 x g (such as a microcentrifuge or a larger floor model) and a water bath incubator that will maintain a temperature of 60-65°C are required for the program. A shaking bacterial incubator is strongly recommended.