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Background Session #6 DNA Sequencing and Using DNA Sequences May 31, 2008. Dr. Simona Bartl Adjunct Professor Moss Landing Marine Labs. Intro to DNA Sequencing. Definition. A technique used to determine the sequence of nucleotide bases in a DNA molecule or fragment. bioweb.wku.edu.
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Background Session #6DNA Sequencing and Using DNA SequencesMay 31, 2008 Dr. Simona Bartl Adjunct Professor Moss Landing Marine Labs
Intro to DNA Sequencing May 31st Workshop
Definition • A technique used to determine the sequence of nucleotide bases in a DNA molecule or fragment bioweb.wku.edu May 31st Workshop
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 May 31st Workshop
Non-examples • DNA sequencing is NOT commonly used for… • DNA fingerprinting • Making monoclonal antibodies • Conducting stem cell research May 31st Workshop
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 May 31st Workshop
Dye-terminator sequencing May 31st Workshop
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) May 31st Workshop