Sanger sequencing

Sanger Sequencing

Sanger sequencing, also known as the chain termination method, is a method of DNA sequencing that was developed by Frederick Sanger and his colleagues in 1977.

Pronunciation

Sanger sequencing: /ˈsæŋ.ər ˈsiː.kwən.sɪŋ/

Etymology

The term "Sanger sequencing" is named after its developer, British biochemist Frederick Sanger. The term "sequencing" comes from the Latin word "sequentia", meaning "following".

Definition

Sanger sequencing is a method of DNA sequencing based on the selective incorporation of chain-terminating dideoxynucleotides by DNA polymerase during in vitro DNA replication. It is a simple and efficient method for sequencing DNA and has been the most widely used DNA sequencing method for over 40 years.

Process

The process of Sanger sequencing begins with a single-stranded DNA template, a DNA primer, DNA polymerase, normal deoxynucleotides (dNTPs), and dideoxynucleotides (ddNTPs). The ddNTPs are the key to the sequencing process as they stop the DNA synthesis due to the lack of a 3' OH group. This results in DNA fragments of varying lengths, which can then be separated by size using electrophoresis. The sequence of the DNA can then be read by the order of the fragments.

Related Terms

• DNA sequencing: The process of determining the precise order of nucleotides within a DNA molecule.
• Dideoxynucleotides: A type of nucleotide used in Sanger sequencing to terminate DNA synthesis.
• DNA polymerase: An enzyme that synthesizes DNA molecules from deoxyribonucleotides.
• Electrophoresis: A laboratory method used to separate mixtures of DNA, RNA, or proteins according to their size.

See Also

• Next-generation sequencing
• Pyrosequencing
• Shotgun sequencing

External links

• Medical encyclopedia article on Sanger sequencing
• Wikipedia's article - Sanger sequencing

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