Download - 5. DNA Sequencing
pp 379 - 381
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