1. 5. DNA Sequencing pp

2. Fredrick Sanger Developed dideoxy termination sequencing method
Nobel Laureate, 1980
Nobel Laureate, 1958; determined insulin sequence

3. DNA Sequencing
The components of a sequencing reaction are:

4. dideoxyribose when both oxygen atoms removed
deoxyribose when oxygen removed
3’ OH functional group necessary for DNA elongation

5. DNA Sequencing
Four reactions are set up; one for each possible nucleotide. A G T C
15% dATP
25% dATP
25% dGTP
15% dGTP
25% dTTP
15% dTTP
25% dCTP
15% dCTP
10% ddATP
10% ddGTP
10% ddTTP
10% ddCTP

6. DNA Sequencing
components in one test tube

7. DNA Sequencing
DNA Sequencing Video

8. DNA Sequencing
Contents of each tube is separated by gel electrophoresis
Which type of gel medium is best suited for this method?

9. DNA Sequencing

10. DNA Sequencing

11. DNA Sequencing