discovery of single nucleotide polymorphisms and mutations by pyrosequencing发现通过焦磷酸测序获得的单核苷酸多态性与变异.pdfVIP

Comparative and Functional Genomics Comp Funct Genom 2002; 3: 51–56. DOI: 10.1002 / cfg.132 Review Discovery of single nucleotide polymorphisms and mutations by Pyrosequencing 1 1,2 Mostafa Ronaghi * and Elahe Elahi 1Stanford Genome Technology Center 855 California Ave. Palo Alto, CA 94304, USA 2 Faculty of Science, University of Tehran, Tehran, Iran *Correspondence to: Abstract Stanford Genome Technology Center, Stanford University, 855 Comparative genomics, analyzing variation among individual genomes, is an area of California Avenue, Palo Alto CA intense investigation. DNA sequencing is usually employed to look for polymorphisms and 94304, USA. mutations. Pyrosequencing, a real-time DNA sequencing method, is emerging as a popular E-mail: mostafa@ platform for comparative genomics. Here we review the use of this technology for mutation scanning, polymorphism discovery and chemical haplotyping. We describe the methodology Received: 23 October 2001 and accuracy of this technique and discuss how to reduce the cost for large-scale analysis. Accepted: 27 November 2001 Copyright # 2002 John Wiley Sons, Ltd. Published online: Keywords: SNP; Pyrosequencing; re-sequencing; bioluminescence; haplotyping; mutation 11 January 2002 detection; luciferase Determination of mutations and polymorphisms in structures [19], mutation detection [2], cDNA analy- a genome is one of the most important tasks in sis [12,14,18], re-sequencing of disease-associated the studies of biological systems today. Three genes [3,8], bacterial typing [11], viral typing [9] DNA sequencing platforms are now being used to