One-Step One-Lane Chemical Sequencing of DNA
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Last Update: 2021-03-01
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Source: Internet
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Author: User
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There are several chemical methods for sequencing
DNA
. The classical method, introduced in 1977 by Maxam and Gilbert (
1
), uses four different chemical reactions to attack, modify, and open the heterocyclic bases at guanine (G), adenine and guanine (A + G), thymine and cytosine (T + C), and cytosine (C) in a radioactive end-labeled DNA fragment. The reactions result also in breakage of the glycosidic bond. A second reaction with hot piperidine is required to cleave the DNA backbone at these positions. The reaction products are thus analyzed by electrophoresis in four different lanes of a polyacrylamide gel. Several alternatives to the basic protocol were developed, as reviewed in (
2
).
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