Pyrosequencing

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Pyrosequencing (also known as 454-sequencing) is a method of DNA Sequencing. It involves using an enzyme called luciferase, amongst others, to determine the nucleotide sequence based upon the light emitted when a correct dNTP binds to a template.

The 454 method of sequencing has increased the speed and efficiency of DNA sequencing. It incorporates a laminar flow method of delivering reagents and washing away unused reagents along with etched fibre optic slides to help measure the light output of the reactions occurring. It is a massively parallel process reducing the cost of sequencing greatly. 

454 sequencing has been shown to give "99.5% accuracy at 200 bases" and is capable of giving ~20 megabases of 110 base-reads taking around 8 hours and looks to increase as the system becomes more refined.

The downsides are that the system still has a relatively low throughput. This increases the cost per base of the system [1].   

Alternatives to this system include the Solexa system [2].

References

  1. Rothberg, J.M. & Leamon, J.H., 2008. The development and impact of 454 sequencing. Nature biotechnology, 26(10), pp.1117-24. Available at: http://www.ncbi.nlm.nih.gov/pubmed/18846085.
  2. Ronaghi M. Pyrosequencing Sheds Light On DNA Sequencing, Genome Res. 2001 11: 3-11


 

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