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Transcript of DNA Sequencing
by Emmett Sherman, Alice Hawkes, and Helen Fiora
DNA sequencing is the comprehensive mapping of the sequence of base pairs in a specific genome.
DNA sequencing was first begun by Fredrick Sanger in 1972, using viral DNA.
The first organism to have its
DNA sequenced was E.Coli.
The first eukaryotic cell to have
its DNA sequenced was
brewer's yeast (Saccharomyces cerevisiae)
The first multicellular organism to have its DNA sequenced was Caenorhabditis elegans, a small worm.
Developed by Allan Maxam and Walter Gilbert in 1976.
1. A restriction enzyme is used to cut the DNA at a specific sequence.
2. The DNA is tagged with P32, a radioactive isotope.
3. The tagged DNA is treated with restriction enzyme again and run through gel electrophoresis, separating the tagged and untagged ends.
4. The sequence to be determined is purified.
5. The base pairs are separated by length.
6. Radiography is performed.
7. X-ray is read.
1. The DNA is separated into single strands.
2. A primer is attached to one of the single strands. Part of the primer is radioactively labeled so it can be found on a gel.
3. The solution is separated into four tubes, labeled A, T, G, and C, for all of the bases.
4. DNA polymerase and four types of dideoxynucleotides are added to each of the tubes.
In addition, each of the tubes gets a specialized dideoxynucleotide. The G tube gets ddGTP, the T tube gets ddTTP, and so on.
5. The DNA is denatured again.
6. The DNA is then run on a gel and read using the radioactive markers.
Also called the Sanger Method, after Frederick Sanger, who first used it.
DNA Sequencing Companies
23 & Me
International HapMap Project
The Human Genome Project (complete 2003)
National Institutes of Health, National Human Genome Research Institute. (2011). Dna sequencing fact sheet. Retrieved from website: http://www.genome.gov/10001177
Dna sequencing. (n.d.). Retrieved from http://www.dnasequencing.org
(2006, October 06). [Print Photo]. Retrieved from http://upload.wikimedia.org/wikipedia/commons/f/f8/Frederick_Sanger2.jpg
Mahboob, F. (2010). Dna sequencing: Maxam gilbert method. Biotech Articles, Retrieved from http://www.biotecharticles.com/Biotech-Research-Article/DNA-Sequencing-Maxam-Gilbert-Method-285.html
Advances in molecular genetics. In (2001). J. Greenberg (Ed.), BSCS Biology: A Molecular Approach (8 ed., pp. 391-415). Chicago, IL: Everyday Learning Corp.
Canfield, E. (1999). Sanger Method for DNA Sequencing. Retrieved October 31, 2013, from http://www.bio.davidson.edu/Bio111/seq.html
Obenrader, S. (n.d.). The Sanger Method. Retrieved October 31, 2013, from http://www.bio.davidson.edu/courses/molbio/molstudents/spring2003/obenrader/sanger_method_page.htm
(2013, October 29). Cost per Genome [Web Graphic]. Retrieved from http://www.genome.gov/images/content/cost_per_genome_apr.jpg
(2013, October 29). Cost per Raw Megabase of DNA Sequence [Web Graphic]. Retrieved from http://www.genome.gov/images/content/cost_per_megabase_apr.jpg