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CHAPTER 8: NUCLEIC ACID DNA SEQUENCING

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. | CHAPTER 8: NUCLEIC ACID DNA SEQUENCING INTERNATIONAL UNIVERSITY SCHOOL OF BIOTECHNOLOGY BIOCHEMISTRY Learning objectives To understand how DNA is sequenced using the most common technique automated dideoxynucleotide sequencing 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 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. 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 . | CHAPTER 8: NUCLEIC ACID DNA SEQUENCING INTERNATIONAL UNIVERSITY SCHOOL OF BIOTECHNOLOGY BIOCHEMISTRY Learning objectives To understand how DNA is sequenced using the most common technique automated dideoxynucleotide sequencing 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 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. 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. INTRODUCTION Today the sequence of a protein can be determined from DNA analyses in just a few days. Because of the straightforward and repetitive nature of the procedure, the sequencing itself is typically performed in centralized facilities where automated machines carry out the reactions and data analysis. GENOMIC DNA Since it was discovered that DNA is the material in the cell that carries our genetic information, understanding DNA has become a primary focus of genetic research. Our chromosomes, or genome, consist of neatly wound strands of DNA. All living organisms, from bacteria to human beings, contain DNA in each of their cells. Each cell contains the entire genetic code for that organism. GENOMIC DNA DNA consists of just four building blocks, or nucleotides. These four building blocks, known by their .