This one-semester, project-based laboratory manual gives junior/senior level students the opportunity to characterize the enzyme alpha-amylase. As students proceed through the sequenced experiments, they will learn the principles of DNA, RNA, and protein structure by using modern-day laboratory techniques. Genetics, cell biology, and organic chemistry are prerequisites. Table of Contents Examine Enzyme Activity of alpha-Amylase 1 Starch Plate Assay 2 Quantitative Enzyme Assay 3 Factors Affecting Enzyme Function Examine alpha-Amylase Proteins 4 Analysis of Protein Structure Using RasMol 5 Analysis of alpha-Amylase Proteins 5A SDS-PAGE 5B Western Blotting Examine DNA Structure 6 Analysis of DNA Structure Using RasMol 7 Isolation of Chromosomal DNA from Bacillus licheniformis Find the alpha-Amylase Gene 8 PCR Amplification and Labeling of Probe DNA 9 Southern Hybridization 9A Restriction Enzyme Cleavage of Chromosomal DNA 9B Denaturation and Transfer of DNA to a Membrane 9C Southern Hybridization and Detection Clone the alpha-Amylase Gene 10 Cloning the alpha-Amylase Gene 10A Cleavage of Chromosomal DNA 10B Cleavage of Plasmid DNA 10C Ligation of Chromosomal and Plasmid DNA 10D Transformation 10E Identification of alpha-Amylase Clones Analyze alpha-Amylase Clones 11 Verification and Mapping of alpha-Amylase Clones 11A Verification of alpha-Amylase Clones Using PCR 11B Isolation of Plasmid DNA from alpha-Amylase Clones 11C Restriction Cleavage and Mapping of alpha-Amylase Plasmid DNA 11D Preservation of Recombinant Strains 11E Southern Analysis of alpha-Amylase Plasmid DNA 12 Enzyme Activity of alpha-Amylase Clones