Official Course Description:   MCCCD Approval:  06/26/01
BIO246      20016-99999	LAB	1 Credit(s)	3 Period(s)
Cellular and Molecular Biology Laboratory
Introduction to classic, and modern, laboratory techniques in molecular biology and cellular biology. Includes introduction to the use of bacteriology in molecular biology, various methods of studying macromolecules found in eukaryotic and prokaryotic cells, cloning, and purification of DNA. Prerequisites or Corequisites: BIO245.

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MCCCD Official Course Competencies:
BIO246   20016-99999	Cellular and Molecular Biology Laboratory

1.	Gather data, analyze data, and present data. (I)
2.	Describe the correct use and application of basic equipment found in a typical molecular or cellular biology laboratory. (I, II, XV)
3.	Demonstrate proficiency in bacteriological technique. (I, IX)
4.	Describe Beer's Law and Spectrophotometry. (II)
5.	Demonstrate proficiency with a light microscope. (II)
6.	Describe the Gram stain (II)
7.	Demonstrate proficiency in protein and DNA purification (III)
8.	Purify both chromosomal and plasmid DNA. (III, VI, VIII, X)
9.	Define the central dogma of molecular biology. (IV)
10.	Demonstrate proficiency in transfomation of bacteria and yeast. (IV, VII, IX)
11.	Describe recombinant DNA techniques. (IV, VIII, IX)
12.	Clone a segment of DNA (e.g., a gene) into a plasmid or expression vector. (IV, IX)
13.	Measure and describe growth kinetics of a eukaryotic organism (yeast). (V)
14.	Describe allelic frequencies and the Hardy-Weinberg Equilibrium Theory (VI)
15.	Develop a protocol, using genetic transformation techniques, that will "correct" genetic deficiencies. (VII)
16.	Confirm identity of a DNA sample through restriction enzyme digestion and mapping. (VIII, XI)
17.	Describe and perform nucleic acid hybridizations (southern blotting). (XII)
18.	Describe PCR and design PCR primer. (XIII, XIV)
19.	Describe and perform maintenance of cultured mammalian cells. (XV)
20.	Describe the correct use of equipment found in a cell culture laboratory (XV)

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MCCCD Official Course Outline:
BIO246   20016-99999	Cellular and Molecular Biology Laboratory

I. Introduction to Data Gathering and Analysis A. Experimental models: membrane permeability and enzyme kinetics B. How to write a lab report C. Basic equipment and bacteriological technique 1. Pipettors 2. Streaking 3. Inoculation 4. Sterile technique II. Spectrophotometry and Light Microscopy A. The Gram stain B. Beer's Law C. Extinction coefficients D. absorption spectra E. Spectrophotometric analysis of DNA III. Size Separation of Biomolecules A. Protein purification B. Chromatography C. DNA purification D. Electrophoresis IV. Cloning and Expressing a Recombinant Protein A. Transformation and its role in molecular biology B. Calculation of transformation efficiency C. The central dogma of molecular biology V. Growth Kinetics of Saccharomyces cerevisea (yeast) VI. PCR amplification of DNA A. The Polymerase Chain Reaction B. Purification and amplification of DNA from student "cheek cells" C. Allelic frequencies D. Application of the Hardy-Weinberg equilibrium theory and population genetics VII. Genetic Transformation of a Eukaryote A. Adenine auxotrophic mutant strains of Saccharomyces cerevisea transformed B. Mutation corrected VIII. Restriction Endonuclease Digestion A. Isolation of chromosomal DNA from an aquatic organism: Vibrio fischeri B. Restriction digestion of Vibrio fischeri Genomic DNA C. Vectors and cloning IX. Gene Cloning: Recombinant DNA A. Creating a Recombinant: "Cloning the "bioluminescence gene" (luxA) from Vibrio fischeri X. DNA Purfication of Bioluminescent Clones XI. Restriction Mapping of Bioluminescent Clones XII. Southern Blotting: Detecting the luxA Gene by Hybridization XIII. PCR (Polymerase Chain Reaction): Detecting the luxA Gene by Amplification XIV. The National Center for Biotechnology Web site: Designing PCR primers for specific regions of the CRHB2 gene of the fern Ceratopteris richardii. XV. Cells in Culture A. The American Type Culture Collection (ATCC) B. Growth Media and Conditions: An introduction C. Cell culture equipment and supplies D. Maintaining cells in culture

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