Multigene amplification and massively parallel sequencing for cancer mutation discovery.

摘要: We have developed a procedure for massively parallel resequencing of multiple human genes by combining highly multiplexed and target-specific amplification process with high-throughput sequencing technology. The is based on oligonucleotide constructs, called selectors, that guide the circularization specific DNA target regions. Subsequently, circularized sequences are amplified in multiplex analyzed using sequencing-by-synthesis As proof-of-concept study, we demonstrate 10 cancer covering 177 exons average sequence coverage per sample 93%. Seven cell lines one normal genomic were studied mutations polymorphisms identified among genes. Mutations TP53 gene confirmed traditional sequencing.