The genomics revolution that reached its climax in 2000 owes its very existence to two men. The first is Frederick Sanger who in 1977 developed the method for DNA sequencing that now bears his name. The second is Leroy Hood, who (with colleagues Michael Hunkapiller and Lloyd Smith) in 1986 took Sanger's method and made it better.

Sanger's enzymatic approach relies on specially modified reagents (2',3'-dideoxynucleotide triphosphates) whose incorporation into a growing DNA strand terminates the extension reaction (see related story, p. 44). The method calls for extending a primer-template pair in the presence of a radioactive marker and, in four parallel reactions, either dideoxy-A, dideoxy-C, dideoxy-G, or dideoxy-T. The resulting products can then be resolved on a high-resolution polyacry-lamide gel to produce a four-lane-wide "ladder" that reveals the template's sequence. Brilliantly inventive, the technique is also painfully laborious, producing a few hundred or perhaps a thousand bases at a...

Article Extras


Figure(PDF, 1.5MB)

Related Article

BioBusiness | DNA Sequencing Industry Sets its Sights on the Future

1 The polymer pump loads the capillaries with separation matrix.

2 The DNA samples are loaded into the array by a short burst of electrophoresis called "electrokinetic injection."

3 The capillary array is then immersed in running buffer.

4 DNA fragments migrate through the capillaries by size, smallest to largest.

5 As they reach the detection window, the laser beam excites the dye molecules, causing them to fluoresce. Light from all 96 capillaries is collected at once, spectrally separated, and focused onto a CCD camera.

6 Computer software interprets the data to produce a graph of intensity versus run time, or an "electropherogram."

Interested in reading more?

Magaizne Cover

Become a Member of

Receive full access to digital editions of The Scientist, as well as TS Digest, feature stories, more than 35 years of archives, and much more!
Already a member?