Pyrosequencing: Difference between revisions
mNo edit summary |
No edit summary |
||
| Line 1: | Line 1: | ||
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 emitted when a correct [[DNTP|dNTP]] binds to a template. | 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 emitted when a correct [[DNTP|dNTP]] binds to a template. | ||
<br> | 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.<ref>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.</ref> | |||
Alternatives to this system include the [[Solexa|Solexa]] system. | |||
<ref name="null">Ronaghi M. Pyrosequencing Sheds Light On DNA Sequencing, Genome Res. 2001 11: 3-11</ref> <references /> | |||
<br> | |||
| | ||
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.
- ↑ 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.
- ↑ Ronaghi M. Pyrosequencing Sheds Light On DNA Sequencing, Genome Res. 2001 11: 3-11