High-throughput pyrosequencing for analysis of single-nucleotide polymorphisms

Mostafa Ronaghi

doi:10.1117/12.472098

21 June 2002 High-throughput pyrosequencing for analysis of single-nucleotide polymorphisms

Mostafa Ronaghi

Author Affiliations +

Proceedings Volume 4626, Biomedical Nanotechnology Architectures and Applications; (2002) https://doi.org/10.1117/12.472098
Event: International Symposium on Biomedical Optics, 2002, San Jose, CA, United States

Abstract

Pyrosequencing is a DNA sequencing technique that takes advantage of the cooperativity of four enzymes in a single- tube to determine the nucleotide composition of a DNA fragment in real-time. In this manuscript we describe the methodology and the use of this technology for analysis of single nucleotide polymorphisms, although, this technique has also been used for sequence determination of difficult secondary structures, mutation detection, EST sequencing, virus and bacteria typing, and re-sequencing of disease genes. Recent break-through has enabled long read data up to 200 nucleotides to be obtained in a single run. Automated microtiter plate based Pyrosequencing systems have been developed allowing DNA analyses of between 5000 to 50,000 samples per day. We are now miniaturizing this technique to reduce the cost for sequencing by at least two order of magnitudes. The array format proofs the feasibility of this system for DNA sequencing.

Citation Download Citation

Mostafa Ronaghi, "High-throughput pyrosequencing for analysis of single-nucleotide polymorphisms", Proc. SPIE 4626, Biomedical Nanotechnology Architectures and Applications, (21 June 2002); doi: 10.1117/12.472098; https://doi.org/10.1117/12.472098

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