A molecular inversion probe assay for detecting alternative splicing.
BMC Genomics
; 11: 712, 2010 Dec 17.
Article
em En
| MEDLINE
| ID: mdl-21167051
ABSTRACT
BACKGROUND:
A sensitive, high-throughput method for monitoring pre-mRNA splicing on a genomic scale is needed to understand the spectrum of alternatively spliced mRNA in human cells.RESULTS:
We adapted Molecular Inversion Probes (MIPs), a padlock-probe based technology, for the multiplexed capture and quantitation of individual splice events in human tissues. Individual MIP capture probes can be quantified using either DNA microarrays or high-throughput sequencing, which permits independent assessment of each spliced junction. Using our methodology we successfully identified 100% of our positive controls and showed that there is a strong correlation between the data from our alternative splicing MIP (asMIP) assay and quantitative PCR.CONCLUSION:
The asMIP assay provides a sensitive, accurate and multiplexed means for measuring pre-mRNA splicing. Fully optimized, we estimate that the assay could accommodate a throughput of greater than 20,000 splice junctions in a single reaction. This would represent a significant improvement over existing technologies.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Sondas Moleculares
/
Técnicas de Sonda Molecular
/
Processamento Alternativo
Idioma:
En
Ano de publicação:
2010
Tipo de documento:
Article