1. Introduction, Chemistry, Protocols & Troubleshooting
Basics Of
Introduction, Chemistry, Protocols & Troubleshooting
DNA
Sequencing
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2. Introduction to: Fluorescent Sequencing and Electrophoresis
Sanger Sequencing
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3. Sanger Sequencing Basic idea
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4. Sanger Sequencing ChemistryNH 2 O HO P
DEOXYRIBONUCLEIC
ACID
(dNTP) N NH 2 O P
DIDEOXYRIBONUCLEIC
ACID
(ddNTP)
terminator HO
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5. Sanger Sequencing Chemistry
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6. Sanger Sequencing Basic idea w/ fluorescence
dNTP
ddNTP
primer
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7. Chemistry overview Dye terminators
ddATP
ddCTP
ddGTP
ddTTP A AC
ACG
ACGT +
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8. Sanger Sequencing Basic idea w/ fluorescence
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9. Sanger Sequencing Cycle sequencing
denaturation
annealing
rxn. mixture
extension
A C G T G
A C G T
labeled products
A C G T A C G T T A
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10. Sanger Sequencing Cycle sequencing steps
10” 4’ 4’ 5”
4oC 8
x 25
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12. Molecules Separation Electrophoresis principle0’
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13. Molecules separation Electrophoresis principle
30’
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14. Molecules separation Electrophoresis principle
45’
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15. Molecules separation Electrophoresis principle
90’
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16. Sanger Sequencing Electrophoresis principle
denaturation
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17. Sanger Sequencing Chemistry
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18. Sequencing Product
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19. Dr. Mohamed Abdel Fattah Application Specialist Molecular Biology AnalysisAB Co. Cairo, Egypt
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