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Basics Of Introduction, Chemistry, Protocols & Troubleshooting

Explore the basics of Sanger sequencing chemistry, fluorescence principles, and electrophoresis techniques. Delve into dye terminators, ddNTPs, and cycle sequencing with detailed steps for sequencing success. Learn the fundamentals from Dr. Mohamed Abdel Fattah, a specialized molecular biologist based in Cairo, Egypt.

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Basics Of Introduction, Chemistry, Protocols & Troubleshooting

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  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

  11. 0’ Molecules Separation Electrophoresis principle

  12. 30’ Molecules separation Electrophoresis principle

  13. 45’ Molecules separation Electrophoresis principle

  14. 90’ Molecules separation Electrophoresis principle

  15. denaturation Sanger Sequencing Electrophoresis principle

  16. Sanger Sequencing Chemistry

  17. Sequencing Product

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

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