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CHAPTER 8: NUCLEIC ACID DNA
SEQUENCING
INTERNATIONAL UNIVERSITY
SCHOOL OF BIOTECHNOLOGY
BIOCHEMISTRY
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Learning objectives
To understand how DNA is sequenced using
the most common technique automated
dideoxynucleotide sequencing
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Content
1. INTRODUCTION
2. GENOMIC DNA
3. DNA PREPARATION
4. SEQUENCING REACTION
5. STRAND SEPARATION
6. PRIMER ANNEALING
7. PRIMER EXTENSION
8. CHAIN TERMINATION
9. PUTTING IT ALL TOGETHER
10. CAPILLARY ELECTROPHORESIS
11. COMPUTER ANALYSIS
12. PROCEDURE SUMMARY
13. CONCLUSION
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INTRODUCTION
Automated DNA sequencing is a core
research tool used by almost every research
biochemistry lab. It is used to determine the
sequence of DNA, or the genetic code, that
serves as the blueprint of life for every
organism on Earth.
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INTRODUCTION
Nucleic acid sequencing is a relatively late arrival
for the sequencing of biological macromolecules
—for up until the late 1980s protein sequencing
was the primary tool for obtaining coding
information found in the molecules of life. Protein
sequencing is a slow and expensive endeavor,
and it could easily take a year or more to
sequence a protein of 500 amino acids.