1. A program of ITEST (Information Technology Experiences for Students and Teachers) funded by the National Science Foundation Background Session #6 DNA Sequencing and Using DNA Sequences May 31, 2008 Dr. Simona Bartl Adjunct Professor Moss Landing Marine Labs

2. May 31st Workshop2 Intro to DNA Sequencing

3. May 31st Workshop3 Definition  A technique used to determine the sequence of nucleotide bases in a DNA molecule or fragment bioweb.wku.edu

4. May 31st Workshop4 Application Examples  DNA sequencing is commonly used to…  Identify useful regions of DNA for other applications  Identify nucleotides in a gene and the inferred protein sequence  Generate data for taxonomic analysis  Identify gene defects associated with certain diseases  Analyze whole genomes

5. May 31st Workshop5 Non-examples  DNA sequencing is NOT commonly used for…  DNA fingerprinting  Making monoclonal antibodies  Conducting stem cell research

6. May 31st Workshop6 Techniques  Maxam-Gilbert (chemical cleavage)  DNA broken into fragments of various lengths  Sanger (chain termination or dideoxy sequencing)  DNA synthesis is blocked at varying sites  Dye-terminator sequencing  Combines Sanger and PCR  Reactions are denatured and separated by size

7. May 31st Workshop7 Dye-terminator sequencing

8. May 31st Workshop8 Using DNA sequences  Finding the right level of variation to differentiate:  Individuals  Populations  Species  Higher taxa  Using levels of variation to determine:  Degrees of relatedness  Time of divergence (based on mutation rate)