Pyrosequencing

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Pyrosequencing (also known as 454-sequencing) is a method of [[DNA Sequencing|DNA Sequencing]]. It involves using an [[Enzyme|enzyme]] called [[Luciferase|luciferase]], amongst others, to determine the [[Nucleotide|nucleotide]] sequence based upon the light&nbsp;emitted when a correct [[DNTP|dNTP]] binds to a template. <ref name="null">Ronaghi M. Pyrosequencing Sheds Light On DNA Sequencing, Genome Res. 2001 11: 3-11</ref> <references />
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Pyrosequencing (also known as 454-sequencing) is a method of [[DNA Sequencing|DNA Sequencing]]. It involves using an [[Enzyme|enzyme]] called [[Luciferase|luciferase]], amongst others, to determine the [[Nucleotide|nucleotide]] sequence based upon the light&nbsp;emitted when a correct [[DNTP|dNTP]] binds to a template.
  
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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.&nbsp;
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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.
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The downsides are that the system still has a relatively low throughput. This increases the cost per base of the system.<ref>Rothberg, J.M. &amp; 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.</ref>&nbsp; &nbsp;
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Alternatives to this system include the [[Solexa|Solexa]]&nbsp;system.
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<ref name="null">Ronaghi M. Pyrosequencing Sheds Light On DNA Sequencing, Genome Res. 2001 11: 3-11</ref> <references />
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Revision as of 22:25, 22 November 2010

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]
  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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