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Intro_W2010 - TABLE OF CONTENTS Page Introduction ii First...

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TABLE OF CONTENTS Page Introduction ii First Lab Period Introduction to the Laboratory 0-1 Experiment 1 Buffers and pH 1-1 Experiment 2 Spectrophotometric Methods: Protein Determination 2-1 Experiment 3 Enzymes: Assays and Kinetics 3-1 Experiment 4 Purification of an Enzyme 4-1 Experiment 5 Electrophoresis of Proteins in Polyacrylamide Gels 5-1 Experiment 6 Human Intron-less Gene of Lactate Dehydrogenase 6-1 Experiment 7 Recombinant DNA 7-1 Experiment 8 Internet Proteomics Tools 8-1 Appendix I Practice Problems AI-1 Appendix II Practice Problems Answers AII-1
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ii INTRODUCTION Overall Course Goals Molecular and Cellular Biology 120L is a laboratory course that stresses analytical and quantitative biochemical techniques. The laboratory skills that make up the course are currently in use in academic, government and biotechnology research institutions throughout the world. For this reason, successful mastery of the techniques taught in this course should prepare you for a wide variety of careers in biology. Standard Laboratory Procedures 1. Read the entire experiment before the beginning of the first laboratory period . Each laboratory session will be very demanding due to the number and complexity of the experiments. As a result, it is essential that you come to the laboratory familiar with the purposes and procedural details of the day's experiments. This means protocols ( i.e., detailed plans for laboratory work), flowsheets, and calculations should all be done in advance. For example, when the dilution of stock solutions is required, you should plan, before coming to the laboratory, how many milliliters of stock are to be diluted to what volume, in order to give the volume and concentration desired for any given procedure. 2. In general, you will work in pairs (or in groups of three). Partners will be changed twice during the quarter. All group members must be aware of the purpose and progress of each phase of the work. Work responsibilities should shift so that all students have an opportunity to practice and learn every technique and procedure. While it is possible for one member of a group to not strongly participate in the laboratory, this would be a mistake since examinations will be based on a solid grasp of the procedures, calculations, potentials and limits of methods and instruments, etc.--- information easily obtained through practice, but hard to get otherwise. 3. During the first part of the first laboratory period, you should familiarize yourself with the types and locations of glassware and instruments, the front bench where solutions and supplies for the day's experiment are stored, the soaking tubs for large glassware, test tubes, pipets, etc. 4. Each student is responsible for the proper use and cleanliness of instruments, equipment and bench space.
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