Sequence Determination

Methods to determine the sequences of bases in DNA were pioneered in the 1970s by Frederick Sanger and Walter Gilbert, whose efforts won them a Nobel Prize in 1980. The Gilbert-Maxam method relies on the different chemical reactivities of the bases, while the Sanger method is based on enzymatic synthesis of DNA in vitro. Both methods measure the distance from a fixed point on DNA to each occurrence of a particular base—A, C, G, or T. DNA fragments obtained from a series of reactions are separated according to length in four “lanes” by gel electrophoresis. Each lane corresponds to a unique base, and the sequence is read directly from the gel. The Sanger method has now been automated using fluorescent dyes to label the DNA, and a single machine can produce tens of thousands of DNA base sequences in a single run.