Sequencing DNA: Principle Create a nested sequence of fragments that can be seperated using gel electrophoresis to determine the sequence of nucleotides.

• Maxim and Gilbert degradation techniques using mutagens.
• Seperate on gels
• Reconstructing the sequence from the nested fragments
• A real autoradiograph.
• Sanger creates a nested set of fragments using dideoxy nucleotide phosphates.
• Using DNA replicating enzymes anComd dideoxy nucleotides we can get a nice series of nested fragments.
• A real Sanger dideoxy autoradiograph.
• Automated Sequencer Printout.
• Why are fluorescing dyes important to this process?
  • Dual labeling experiments - what does this mean to you.
• Computer scanning for ORFs(open reading frames)
• Archibald Garrod - 1902 - Alkaptonuria

This web site is provided for instruction in Botany and Zoology 342

by Kenneth G. Wilson,
Professor of Botany
Miami University
wilsonkg@muohio.edu