Multigene amplification and massively parallel sequencing for cancer mutation discovery
2007 (English)In: Proceedings of the National Academy of Sciences of the United States of America, ISSN 0027-8424, E-ISSN 1091-6490, Vol. 104, no 22, 9387-9392 p.Article in journal (Refereed) Published
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.
Place, publisher, year, edition, pages
2007. Vol. 104, no 22, 9387-9392 p.
cancer analysis, high-throughput sequencing, multiplex amplification
Medical and Health Sciences
IdentifiersURN: urn:nbn:se:uu:diva-101036DOI: 10.1073/pnas.0702165104ISI: 000246935700055PubMedID: 17517648OAI: oai:DiVA.org:uu-101036DiVA: diva2:211596