Biotechnology Proteins to PCR A Course in Strategies and Lab Techniques [Paperback]

1. Introduction to the Biotechnology Laboratory.- 1.1Overview.- 1.2 Background.- Scope of Biotechnology.- Working in the Biotech-nology Laboratory.- Laboratory Safety.- Notes, Records, and Labels.- Housekeeping.- Preparation and Storage of Buffers.- Preparation and Storage of Media.- Care and Maintenance of Cultures.- General Rules for Protein Handling.- Storage of Nucleic Acids.- 1.3 Experimental Design and Procedures.- Preparation ofYPD and YPG Media.- Streak Plating of Escherichia coli and Saccharomyces carlsbergensis.- Microscopic Observation of Bacteria and Yeast.- Inoculation of YPD and YPG with Saccharomyces carlsbergensis.- Study Questions.- Further Readings.- 2. Introduction to Proteins.- 2.1 Overview.- 2.2 Background.- Protein Function.- Characterization of Proteins.- Measurement of Biological Activity.- 2.3 Experimental Design and Procedures.- Time Course Study of ?-Galactosidase Assay.- Analysis of Culture Broth for a-Galactosidase Activity by the Standard a-Gal Enzyme Assay.- Colorimetric Protein Assays.- Biuret Protein Assay.- Lowry Protein Assay.- Bradford Protein Assay.- Preparation and Inoculation of YP Broth for ?-Galactosidase Production.- Study Questions.- Further Readings.- 3. Protein Isolation and Preparation of Crude Extract.- 3.1 Overview.- 3.2 Background.- Selection of Protein Source.- Cell Disruption Techniques.- Factors Affecting Activity in the Crude Extract.- 3.3 Experimental Design and Procedures.- ?-Gal Assay of Yeast Culture Broth.- ?-Gal Assay of the Yeast Supernatant.- Preparation of Yeast Protoplasts.- Lysis of the Yeast Protoplast Cells.- Collection of Supernatant and Determination of Total Protein.- Stability Study of Yeast Supernatant.- Study Questions.- Further Readings.- 4. Batch Purification of Proteins.- 4.1 Overview.- 4.2 Background.- Bulk Precipitation Techniques.- Batch Protein Capture.- Gel Binding Capacity.- 4.3 Experimental Design and Procedures.- Determination of Binding pH.- Capture of ?-Galactosidase by Batch Ion ExlÓß