Basics Of Introduction, Chemistry, Protocols & Troubleshooting

1. Basics Of Introduction, Chemistry, Protocols & Troubleshooting DNA Sequencing

2. Introduction to:Fluorescent Sequencing and Electrophoresis Sanger Sequencing

3. Sanger Sequencing Basic idea

4. NH NH 2 2 O O O P O O P O N N O O O N O N O O HO HO DIDEOXYRIBONUCLEIC ACID (ddNTP) terminator DEOXYRIBONUCLEIC ACID (dNTP) Sanger Sequencing Chemistry

5. Sanger Sequencing Chemistry

6. Sanger Sequencing Basic idea w/ fluorescence dNTP ddNTP primer

7. ddATP A ddCTP AC ddGTP ACG ddTTP ACGT Chemistry overviewDye terminators +

8. Sanger Sequencing Basic idea w/ fluorescence

9. Sanger Sequencing Cycle sequencing denaturation annealing rxn. mixture extension A CG T G A CG T A CG T labeled products A C G T T A A

10. Sanger Sequencing Cycle sequencing steps 50°C 50°C 96°C 60°C 10” 4’ 4’ 5” 4oC 8 x 25

12. 0’ Molecules Separation Electrophoresis principle

13. 30’ Molecules separation Electrophoresis principle

14. 45’ Molecules separation Electrophoresis principle

15. 90’ Molecules separation Electrophoresis principle

16. denaturation Sanger Sequencing Electrophoresis principle

17. Sanger Sequencing Chemistry

18. Sequencing Product

19. Dr. Mohamed Abdel FattahApplication SpecialistMolecular BiologyAnalysisAB Co.Cairo, Egypt