CHEM 373 Experimental Biochemistry

This lab course is highly recommended to enhance the introductory biochemistry course and as preparation for further study of biochemistry and related fields.  The course introduces students to the process and techniques necessary to build research based biochemical models.  Procedures (see links below) include work in characterization and isolation of proteins and manipulation of DNA.  Particular emphasis is based on understanding the functional attributes of isolated biological molecules using enzyme kinetics.

Dr. Muth has been developing this course since the Spring of 2003 using cystathionine-beta lyase as the enzyme of interest.  He feels that the laboratory curriculum could be adapted to study almost any enzyme that has been cloned into an expression vector.

A recent update was presented at the "Interdisciplinary Science Education: Institutional Examples, Lessons Learned and Challenges" conference at the Midstates Consortium for Mathematics and Science sponsored by the Pew Charitable Trust.

A Research-Style Biochemistry Lab: Collaborating on the Integration of Research and Teaching at Two Institutions


Weekly Schedule of Experiments

Week 1            Design: Introduction, Overview and Chemical Mechanism of CBL (overview)

Week 2            Design: Molecular Modeling and Mutant Design (Pre-lab 1 due, overview)

Week 3            DNA: PCR, ligation and transformation (Pre-lab 2 due)

Week 4            DNA: Plasmid isolation analysis (Pre-lab 3 due, Analysis assignment 1 due)

Week 5            DNA: Sequencing (Pre-lab 4 due, Analysis assignment 2 due)

Week 6            Protein: Isolation and purification (Pre-lab 5 due)

Week 7*            Protein: Concentration determination (Pre-lab 6 due, Analysis assignment 3 due)

Week 8            No Lab

Week 9*            Protein: Molecular weight and structure analysis (Pre-lab 7 due, Analysis assignment 4 due)

Week 10            Protein: CBL kinetic assay optimization (Pre-lab 8 due, Analysis assignment 5 due, overview)

Week 11          CBL kinetics  (Pre-lab 9 due

Week 12          CBL kinetics (Analysis assignment 6 due)

Week 13          CBL kinetics

Week 14            Presentations

*could be combined and completed in a single week


For experimental details, tips, hints and background please consult the following links:

The Virtual Lab Manual

Appendix I.  Pipettors and pipetting
Appendix II.  Concentration and dilution
Appendix III.  Spectroscopy
Appendix IV.  Gel electrophoresis
Appendix V.  Creating images on the Lumi-Imager F1 device
Appendix VI.  Creating figures


Related Links

The Protein Data Bank
Biopolymer Calculator
NEB WebCutter
NCBI Blast Search
DNA Sequencing
DNA Synthesis
 


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