Sanger sequencing

Sanger sequencing, also known as the chain termination method, is a method of DNA sequencing first commercialized by Frederick Sanger and his colleagues in 1977. This method is based on the selective incorporation of chain-terminating dideoxynucleotides by DNA polymerase during in vitro DNA replication.

History[edit]

The Sanger sequencing method was developed in 1977 by two-time Nobel laureate Frederick Sanger and his colleagues. This method quickly replaced the then-used plus and minus method and became the most widely used DNA sequencing method for the next 40 years.

Method[edit]

The Sanger sequencing method involves four separate DNA synthesis reactions. Each reaction includes a DNA template, a DNA primer, the four deoxyribonucleotide (dNTP) bases, DNA polymerase, and a small amount of one of the four dideoxynucleotide (ddNTP) bases. The ddNTPs cause DNA synthesis to stop, as they lack the 3'-OH group required for the formation of a phosphodiester bond between two nucleotides, causing DNA polymerase to cease extension of DNA.

Applications[edit]

Sanger sequencing is used in a wide range of applications, including gene sequencing, mutation detection, and genetic disease diagnosis. It is also used in research for studying genetic disorders, cancer, and infectious diseases.

Limitations[edit]

While Sanger sequencing is a reliable and accurate method for DNA sequencing, it has several limitations. These include the inability to sequence very long DNA molecules, the requirement for a large amount of purified DNA, and the relatively high cost and time-consuming nature of the method.