Pyrosequencing

Pyrosequencing is a non-electrophoretic DNA sequencing (determining the order of nucleotides in DNA) method based on the "sequencing by synthesis" principle, in which the sequencing is performed by detecting the nucleotide incorporated by a DNA polymerase. Pyrosequencing relies on light detection based on a chain reaction when pyrophosphate is released, hence, the name given it.

Pyrosequencing is a non-electrophoretic DNA sequencing (determining the order of nucleotides in DNA) method based on the "sequencing by synthesis" principle, in which the sequencing is performed by detecting the nucleotide incorporated by a DNA polymerase. Pyrosequencing relies on light detection based on a chain reaction when pyrophosphate is released, hence, the name given it. ==Limitations== Pyrosequencing can have difficulty accurately determining the length of homopolymer regions, which can result in sequencing errors. This can make the process of genome assembly more difficult, particularly for sequences containing a large amount of repetitive DNA. Also, lack of a proof-reading activity limits accuracy of this method. ==Commercialization== Pyrosequencing AB, a company based in Uppsala, Sweden, was founded with venture capital provided by HealthCap in order to commercialize machinery and reagents for sequencing short stretches of DNA using the pyrosequencing technique. Pyrosequencing AB was listed on the Stockholm Stock Exchange in 1999. When Pyrosequencing AB acquired Biotage LLC, a U.S.-based company, and other companies, in 2003, the company was renamed Biotage AB. The pyrosequencing and other biomedical units of Biotage AB were sold to Qiagen in 2008. The pyrosequencing technology was licensed to 454 Life Sciences. 454 developed an array-based pyrosequencing technology that emerged as a platform for large-scale DNA sequencing, including genome sequencing and metagenomics. Roche acquired 454 Life Sciences, and announced the discontinuation of the 454 sequencing platform in 2013. The 454 sequencing platform was replaced, in part, by Illumina dye sequencing, and by Applied Biosystems sequencing products. ==Further reading== For the corresponding PDF document, see this link. ==References==

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