Multigene amplification and massively parallel sequencing for cancer mutation discovery
- PMID: 17517648
- PMCID: PMC1871563
- DOI: 10.1073/pnas.0702165104
Multigene amplification and massively parallel sequencing for cancer mutation discovery
Abstract
We have developed a procedure for massively parallel resequencing of multiple human genes by combining a highly multiplexed and target-specific amplification process with a high-throughput parallel sequencing technology. The amplification process is based on oligonucleotide constructs, called selectors, that guide the circularization of specific DNA target regions. Subsequently, the circularized target sequences are amplified in multiplex and analyzed by using a highly parallel sequencing-by-synthesis technology. As a proof-of-concept study, we demonstrate parallel resequencing of 10 cancer genes covering 177 exons with average sequence coverage per sample of 93%. Seven cancer cell lines and one normal genomic DNA sample were studied with multiple mutations and polymorphisms identified among the 10 genes. Mutations and polymorphisms in the TP53 gene were confirmed by traditional sequencing.
Conflict of interest statement
Conflict of interest statement: M.N. and S.F. are cofounders of Olink AB and hold commercial rights to selector technology, and F.D and J.S. have business agreements with Olink AB.
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References
-
- Paez JG, Janne PA, Lee JC, Tracy S, Greulich H, Gabriel S, Herman P, Kaye FJ, Lindeman N, Boggon TJ, et al. Science. 2004;304:1497–1500. - PubMed
-
- Lynch TJ, Bell DW, Sordella R, Gurubhagavatula S, Okimoto RA, Brannigan BW, Harris PL, Haserlat SM, Supko JG, Haluska FG, et al. N Engl J Med. 2004;350:2129–2139. - PubMed
-
- Sjoblom T, Jones S, Wood LD, Parsons DW, Lin J, Barber TD, Mandelker D, Leary RJ, Ptak J, Silliman N, et al. Science. 2006;314:268–274. - PubMed
-
- Patil N, Berno AJ, Hinds DA, Barrett WA, Doshi JM, Hacker CR, Kautzer CR, Lee DH, Marjoribanks C, McDonough DP, et al. Science. 2001;294:1719–1723. - PubMed
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