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Fredrick SangerDeveloped dideoxy

termination sequencing method

Nobel Laureate, 1980

Nobel Laureate, 1958; determined insulin sequence

DNA SequencingThe components of a sequencing reaction are:1. 2. 3. 4. 5.

deoxyribose when oxygen removed

dideoxyribose when both oxygen atoms removed

3’ OH functional group necessary for DNA elongation

DNA SequencingFour reactions are set up; one for each possible nucleotide.

A G T C

15% dATP 25% dATP 25% dATP 25% dATP

25% dGTP 15% dGTP 25% dGTP 25% dGTP

25% dTTP 25% dTTP 15% dTTP 25% dTTP

25% dCTP 25% dCTP 25% dCTP 15% dCTP

10% ddATP 10% ddGTP 10% ddTTP 10% ddCTP

DNA Sequencing

components in one test tube

DNA Sequencing

DNA Sequencing Video

DNA Sequencing

Contents of each tube is separated by gel electrophoresis

Which type of gel medium is best suited for this method?

DNA Sequencing

DNA Sequencing

DNA Sequencing