Use of a chloroplast rRNA gene to introduce basic molecular biology techniques

Abstract

AbstractWe report a set of experiments that has been used for 14 years in an upper division undergraduate molecular biology laboratory course to introduce basic molecular biology techniques including DNA isolation, gene subcloning, restriction mapping, plant genomic DNA isolation, Southern blot hybridization, and DNA sequence analysis. This set of experiments is very useful for students who have limited or no previous experience with these techniques. A single gene, the 23S rRNA gene from tobacco chloroplast DNA, is used throughout these experiments. This allows for integration of the techniques and illustrates to students a natural progression of experiments to use in analyzing a new gene. The entire sequence of laboratory exercises spans 10–12 laboratory periods of 2.5–3 h each.