Individual Assignment: Dideoxy Chain Termination DNA Sequencing

The Dideoxy Chain-termination method for sequencing DNA, or the Sanger Sequencing
Method, is a technique for sequencing a strand of DNA. DNA sequencing is determining the
exact order of nucleotides in a piece of DNA (Campbell, 2005). It was developed by Frederick
Sanger in 1977 (Sanger method, Cold Spring Harbor). It is the type of DNA sequencing most
frequently used today.
The thing that makes the Sanger method possible is specially altered fluorescent dideoxy
nucleotides. Dideoxy nucleotides are regular nucleotides except with a hydrogen group instead
of a hydroxyl (OH) group (Tech council, Sanger DNA). The hydrogen group prevents further
nucleotides from being added, and so the chain of replication is terminated whenever a dideoxy
nucleotide is added, creating many fragments of DNA. (Tech council, Sanger DNA). The
fluorescence allows the different nucleotides to be determined when they are scanned by a laser.
(DNA sequencing, the Welcome Trust).
During the process of the Sanger method, four reaction tubes are set up. Each tube has
DNA polymerase, the DNA to be sequenced, a full supply of normal nucleotides, and one kind
of dideoxy nucleotide (either A, G, T, or C.) (Micklos, 2003). As the DNA polymerase replicates
the DNA, the dideoxy nucleotides cause it to fall off and stop at varying points along it, creating
many different length fragments in each tube. (Sanger method, Cold Spring Harbor). The
fragments are then pieced together to form a coherent stream of DNA information. (Sanger
method, Cold Spring Harbor).
In order to do this, the fragments must be sorted out by length. The fragments are run
through a gel full of very small capillaries, like a maze. The longer fragments get stuck more,
and take longer to get through the maze, while the shorter ones are faster. This organizes them
from short to longest. They are then scanned by a laser, and recorded as different colored bands.
(DNA Sequencing, the Welcome Trust). These colors stand for each nucleotide, and their order
shows the nucleotide sequence of the DNA molecule.
Sanger won a Nobel Prize for his advancement, and his method has since been used
extensively in the Human Genome Project. His work was and is a major advancement in
understanding the role genes play in our lives and diseases.

References:
Campbell, Reece. Biology. 7th ed. San Francisco: Pearson Education Inc., 2005. Print.
Council, Tech. "The Sanger DNA Sequencing Methods." Tech Council. N.p., n.d. Web. 04 May
2014.
DNA sequencing - the Sanger method. The Welcome Trust. Web. 4 May 2014.
<http://www.wellcome.ac.uk/Education-resources/Education-and-
learning/Resources/Animation/WTDV026689.htm>.
Micklos, David A., Greg A. Freyer, and David A. Crotty. DNA Science. Second ed. Cold Spring
Harbor, New York.: John Inglis, 2003. Print.
Sanger method of DNA sequencing, 3D anim. Cold Spring Harbor Laboratory
Web. 4 May 2014. <http://www.dnalc.org/view/15479-Sanger-method-of-DNA-
sequencing-3D-animation-with-narration.html>.