CR, June 29, 2005

Nucleic Acids Chemistry

Outline of Today's Lecture
 

DNA Sequencing

1.  Comparison with Peptide Sequencing
 

2.  Maxam-Gilbert Method

     -  Base-specific fragmentation

     -  Separation of strands by PAGE
 

3.  Dideoxy- or Chain Terminator Method

     -  2',3'-Dideoxynucleotide triphosphates

     -  Synthesis of truncated complementary strands
 

4.  Automated 1 Reaction/4 Fluorophore Method

     -  Big Dye-Labeled ddNTPs

     -  Read-out by capillary electrophoresis
 

5.  Genome Sequencing
 

References:
    - Section 28-6, Voet & Voet, Biochemistry 2nd edition
    - Section 7-2, Voet & Voet, Biochemistry 3rd edition
 

For a Powerpoint presentation on sequencing (by U. Auckland)
    See:  www.sbs.auckland.ac.nz/info/ schools/biotechnology/Sequencing.ppt