B_+Introduction+F2010 - ii TABLE OF CONTENTS Page...

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TABLE OF CONTENTS Page Introduction ii First Lab Period Introduction to the Laboratory 0-1 Experiment 1 Weak Acid Buffers 1-1 Experiment 2 Spectrophotometric Methods: Protein Determination 2-1 Experiment 3 Enzymes: Assays and Kinetics 3-1 Experiment 4 Purification of Bovine LDH 4-1 Experiment 5 Electrophoresis of Proteins in Polyacrylamide Gels 5-1 Experiment 6 Restriction mapping of a Human Intron-less Gene of LDH 6-1 Experiment 7 Expression of a ORF from a cyanobacterium in an E.coli host 7-1 Experiment 8 Introduction to Internet Proteomics Tools 8-1 Appendix I Study problems A-1 Appendix II Answers to Study Problems A-2 Appendix III Math Camp A-3 ii
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INTRODUCTION Course Goals The laboratory class MCB120L stresses analytical and quantitative biochemical techniques. Laboratory skills taught in this course are in use in public (academia and government institutions), and private (biotechnology institutions) research endeavors worldwide. Consequently, the student should focus on becoming adept at applying the relatively simple mathematical computations, and constructing the graphical and tabular presentations of data, required in this course. Successful mastery of the techniques practiced in this course will prepare the student for a wide variety of careers in the life sciences. Standard Laboratory Procedures 1. READ the ENTIRE experiment before starting each experiment. Typically eight to ten exercises are performed within the ten-week quarter. Each requires data analysis, an interpretation of the results, and a discussion of conclusions in a report. This makes this course very demanding. Each laboratory session should be completed in the scheduled time. You want to proceed through the steps of each session’s tasks with deliberate and competent manipulations of equipment and materials to help ensure good data. Lack of preparation puts incredible pressure on you, and your partners, to finish experiments, which often leads to rushing through experimental steps and acquiring inferior data. 2. Each lab session will typically begin with a pre-lab discussion by your instructor. This is NOT A SUBSTITUTE for being prepared before coming to lab. It is essential that you come to lab familiar with the purpose and procedural details of the experiments. This means that protocols, flow sheets, and pre-lab calculations should be done before coming to lab. For example, if dilutions are required in the experiment, then a set of pertinent calculations and procedural steps should be outlined prior to starting the experiment. These are easily referred to when additional dilutions are required. The pre- lab discussion, then, is to cover pertinent points of an experiment such as safety issues, assembling equipment, which steps are critical to do exactly as written and which are less critical, etc. 3. You will usually work in groups of three; larger groups are sometimes necessary. Each member of
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This note was uploaded on 11/21/2010 for the course MCB 120L 69059 taught by Professor Fairclough during the Fall '10 term at UC Davis.

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B_+Introduction+F2010 - ii TABLE OF CONTENTS Page...

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