Table of Contents

I: BASIC TECHNIQUES.

Labs.
 1. Measurements, Micropipetting, and Sterile Techniques. 2. Basic Culture Techniques. 3. DNA Restriction and Electrophoresis.

II: DNA MODIFICATION.

 4. Effects of DNA Methylation on Restriction.

III: PLASMID TRANSFORMATION AND IDENTIFICATION.

 5. Rapid Colony Transformation of E. coli with Plasmid DNA.
 6. Purification and Identification of Plasmid DNA.

IV: PLASMID RECOMBINATION AND IDENTIFICATION.

 7. Recombination of Antibiotic Resistance Genes.
 8. Transformation of E. coli with Recombinant DNA.
 9. Replicating to Identify Mixed E. coli Populations.
10. Purification and Identification of Recombinant Plasmid DNA.

V: RESTRICTION MAPPING OF LINEAR AND CIRCULAR CHROMOSOMES.

11. Restriction Mapping of the l Chromosome.
12. Restriction Mapping of the Plasmid pBR322.

VI: SOUTHERN HYBRIDIZATION.

13. Southern Hybridization of l DNA.

VII: GENOMIC LIBRARY CONSTRUCTION AND ANALYSIS.

14. Construction of Genomic Library of l DNA.
15. Transformation of E. coli with l Library.
16. Colony Hybridization of l Library.
17. Purification and Identification of l Clones.

VIII: CLONING BY POLYMERASE CHAIN REACTION.

18. Amplification and Purification of l DNA Fragment.
19. Recombination of PCR Product and Plasmid pBLU.
20. Transformation of E. coli with PCR/pBLU Product.
21. The Purification and Identification of PCR/pBLU Recombinants.

IX: HUMAN DNA FINGERPRINTING.

22. Detection of an Alu Insertion Polymorphism by Polymerase Chain Reaction.
23. Detection of a VNTR Polymorphism by Polymerase Chain Reaction.
Appendix I: Equipment, Supplies, and Reagents.
Appendix II: Recipes for Media, Reagents, and Stock Solutions.
Appendix III: Restriction Map Data for pAMP, pKAN, pBLU, l and Bacteriophage.
Appendix IV: Sequence Data for pAMP, pKAN, pBLU, and Bacteriophage l.

About the Author

Mark Bloom earned his Bachelor of Science in biology at Kent State University and his Ph.D. at Rensselaer Polytechnic Institute. Mark is the Assistant Director of the DNA Learning Center at Cold Spring Harbor Laboratory. Currently, Mark coordinates the DNA Science and Advanced Science Workshop Programs, and he also supervises Bio2000, a teaching laboratory. In addition to teaching, developing educational materials, and writing grant proposals, Mark is the principal investigator of training programs for precollege and college faculty and public opinion leaders (funded by the National Science Foundation, Department of Education, Department of Energy, and the Howard Hughes Medical Institute). Mark was the first to develop educational kits that feature polymerase chain reaction. He has seventeen publications to his credit and he was the editor for DNA Science: A First Course in Recombinant DNA Technology (Carolina Biological Supply Company and Cold Spring Harbor Laboratory Press).

Greg Freyer holds two Bachelor of Science degrees; one is in biology from the University of Cincinnati, and the other is in chemistry from Miami University. He also has a Ph.D. in biochemistry from The University of Missouri. Currently, he is an assistant professor at Columbia University. Greg has twelve publications and professional papers to his credit, including DNA Science: A First Course in Recombinant DNA Technology (Carolina Biological Supply Company and Cold Spring Harbor Laboratory Press).

David Micklos holds a Bachelor of Science in biology and a Master of Arts in journalism with an emphasis in science communication, public relations, and research methods. He founded the Cold Spring Harbor Laboratory's precollege education programs in 1985 and the DNA Learning Center in 1987 — the nation's first science center solely devoted to public genetics education. He is responsible for the Center's programs and capital development as well as supervising the staff. Micklos developed the laboratory course DNA Science and played a key role in establishing mobile laboratories for nationwide teacher-training. He is also a principal investigator of training programs for precollege, college faculty, and public opinion leaders (funded by the National Science Foundation, Department of Education, Department of Energy, and the Howard Hughes Medical Institute).