1. REVIEW OF MOLECULAR GENETICS DR. EDELBERG

2. Genes, DNA, & Chromosomes

3. Anatomy of DNA

4. Gene anatomy-simple

5. Anatomy of a Gene

6. DNA LIBRARIES  DNA library - a random collection of DNA fragments from an organism cloned into a vector  Ideally contains at least one copy of every DNA sequence.  Easily maintained in the laboratory  Can be manipulated in various ways to facilitate the isolation of a DNA fragment of interest to a scientist.  Numerous types of libraries exist for various organisms - Genomic and cDNA  DNA library - a random collection of DNA fragments from an organism cloned into a vector  Ideally contains at least one copy of every DNA sequence.  Easily maintained in the laboratory  Can be manipulated in various ways to facilitate the isolation of a DNA fragment of interest to a scientist.  Numerous types of libraries exist for various organisms - Genomic and cDNA

7. Genomic Library

8. Construction of cDNA Library

9. Genomic Vs. cDNA Library Genomic Library Promoters Introns Intergenic Non-expressed genes Genomic Library Promoters Introns Intergenic Non-expressed genes cDNA Library Expressed genes Transcription start sites Open reading frames (ORFs) Splice points cDNA Library Expressed genes Transcription start sites Open reading frames (ORFs) Splice points

10. Extraction of mRNA Collect and grind up animals in mild denaturing solution Spin out debris (Tissue, membranes, etc) Treat with DNAse (removes DNA) Treat with Phenol (removes protein)

11. Synthesis of cDNA from mRNA

12. Cloning DNA into vector

13. Cloning into vector- detailed

14. Cloned into multiple cloning site

15. Plasmid Characteristics FOri FSelectable Marker F3.6 Kb FColor screen FMCS FCircular DNA FOri FSelectable Marker F3.6 Kb FColor screen FMCS FCircular DNA

16. Color Screen

17. Transformation

18. Over night