Sequencing Platform
The sequencing platform offers plasmid or PCR product sequencing using the Sanger technique.
Presentation
The platform primarily works on genes associated with various genetic diseases, mainly those affecting the retina. Exons and flanking intronic regions are amplified by PCR, enzymatically purified (exonuclease I / shrimp alkaline phosphatase), and then sequenced with a sequence mix (BigDye Terminator v1.1 Cycle Sequencing Kit, Applied Biosystems) analyzed on the sequencer.
Results are interpreted using SeqScape software, Applied Biosystems.
The platform validates variants identified by targeted high-throughput sequencing or whole exome sequencing and performs family co-segregation analyses or control subject analyses for selected variants.
The platform also offers plasmid sequencing to control sequences after cloning or cellular overexpression analyses.
Expertise
The sequencing platform of the Institute of Vision specializes in the following areas of expertise:
- Sanger sequencing
- Genes associated with eye diseases
Equipment
- ABI 3730 Genetic Analyzer, Applied Biosystems, 48 capillaries
- SeqScape software, Applied Biosystems