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Introductory Techniques | |
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Introduction to the Laboratory: Basic Equipment and Bacteriological Techniques | |
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Preparation of Media and Reagents Used in Molecular Biology | |
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Isolation of Luminescent Bacteria from Natural Sources | |
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Restriction Digestion and Agarose Gel Electrophoresis of DNA | |
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DNA Isolation and Analysis | |
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Isolation of Chromosomal DNA from Vibrio fischeri | |
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Large-Scale Purification of Plasmid DNA | |
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Spectrophotometric Analysis of DNA | |
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Cloning the Lux Operon | |
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Restriction Digestion of Vibrio fischeri Genomic DNA and Plasmid Vector | |
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Quantification of Genomic DNA by Fluorometry and Agarose Plate Fluorescence | |
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Ligation of Restriction Fragments of Vibrio fischeri DNA to Plasmid Vector | |
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Preparation of Competent Escherichia coli DH5 | |
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Transformation of Competent Escherichia coli DH5 with Recombinant Plasmids | |
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Screening the Vibrio fischeri Genomic Library for Light Producing Clones | |
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Restriction Mapping and Southern Blotting | |
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Small-Scale Plasmid Isolations (Mini-preps) from Bioluminescent Clones | |
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Restriction Mapping of Plasmids from Bioluminescent Clones | |
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Southern Blotting and Hybridization to Detect the luxA Gene | |
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Subcloning the Lux A Gene | |
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Restriction Digestion of lux Plasmids and Cloning Vector for Subcloning luxA | |
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Gel Purification of DNA Restriction Fragments Containing luxA | |
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Subcloning luxA into A Plasmid Vector | |
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Transformation of Competent Escherichia coli DH5 with Subcloned DNA | |
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Colony Hybridization to Screen for luxA Subclones | |
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Small-Scale Plasmid Isolations (Mini-Preps) from luxA Clones | |
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Advanced Techniques | |
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Amplification of luxA from Natural Isolates by the Polymerase Chain Reaction (PCR) | |
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Southern Blotting and Hybridization of PCR Products | |
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DNA Sequencing of lux Genes from Plasmid Templates | |
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Computer Analysis of DNA Sequences Using the World Wide Web | |
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Mapping the Vibrio fischeri Genome by Pulsed Field Gel Electrophoresis | |
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Independent Projects in Molecular Biology | |
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Appendices | |
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The Metric System and Units of Measure | |
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Centrifugation | |
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Spectrophotometry | |
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Agarose and Polyacrylamide Gel Electrophoresis | |
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Methylene Blue Staining of Agarose Gels | |
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Nucleic Acid Hybridization | |
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Alcohol Precipitation of Nucleic Acids | |
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Care and Handling of Enzymes | |
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Restriction Endonucleases | |
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Enzymes Used in Molecular Biology | |
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Maps of Cloning Vectors and Bacteriophage Lambda | |
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Proper Handling and Disposal of Hazardous Materials | |
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Procedures and Precautions for the Use of Radioisotopes | |
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Equipment and Supplies | |
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Media and Reagents | |
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Bacterial Strains | |
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Lists of Suppliers | |
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Recommended References | |
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Restriction Mapping Problems | |