Target enrichment using parallel nanoliter quantitative PCR amplification.
BMC Genomics
; 15: 184, 2014 Mar 10.
Article
em En
| MEDLINE
| ID: mdl-24612714
BACKGROUND: Next generation targeted resequencing is replacing Sanger sequencing at high pace in routine genetic diagnosis. The need for well validated, high quality enrichment platforms to complement the bench-top next generation sequencing devices is high. RESULTS: We used the WaferGen Smartchip platform to perform highly parallelized PCR based target enrichment for a set of known cancer genes in a well characterized set of cancer cell lines from the NCI60 panel. Optimization of PCR assay design and cycling conditions resulted in a high enrichment efficiency. We provide proof of a high mutation rediscovery rate and have included technical replicates to enable SNP calling validation demonstrating the high reproducibility of our enrichment platform. CONCLUSIONS: Here we present our custom developed quantitative PCR based target enrichment platform. Using highly parallel nanoliter singleplex PCR reactions makes this a flexible and efficient platform. The high mutation validation rate shows this platform's promise as a targeted resequencing method for multi-gene routine sequencing diagnostics.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Reação em Cadeia da Polimerase
Limite:
Humans
Idioma:
En
Ano de publicação:
2014
Tipo de documento:
Article