11 Facts About Sanger sequencing

Sanger sequencing is a method of DNA sequencing that involves electrophoresis and is based on the random incorporation of chain-terminating dideoxynucleotides by DNA polymerase during in vitro DNA replication.

However, the Sanger method remains in wide use for smaller-scale projects and for validation of deep sequencing results.

Sanger sequencing is still actively being used in efforts for public health initiatives such as sequencing the spike protein from SARS-CoV-2 as well as for the surveillance of norovirus outbreaks through the Center for Disease Control and Prevention's CaliciNet surveillance network.

DNA sample is divided into four separate Sanger sequencing reactions, containing all four of the standard deoxynucleotides and the DNA polymerase.

Dye-primer Sanger sequencing facilitates reading in an optical system for faster and more economical analysis and automation.

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Dye-terminator Sanger sequencing utilizes labelling of the chain terminator ddNTPs, which permits Sanger sequencing in a single reaction rather than four reactions as in the labelled-primer method.

Sanger sequencing is the "gold standard" for norovirus surveillance methods for the Center for Disease Control and Prevention's CaliciNet network.

In contrast, PCR-based cloning and next-generation Sanger sequencing technologies based on pyroSanger sequencing often avoid using cloning vectors.

Microfluidic Sanger sequencing is a lab-on-a-chip application for DNA sequencing, in which the Sanger sequencing steps are integrated on a wafer-scale chip using nanoliter-scale sample volumes.

Sanger sequencing methods achieve maximum read lengths of approximately 800bp .

Ultimate goal of high-throughput Sanger sequencing is to develop systems that are low-cost, and extremely efficient at obtaining extended read lengths.