The sequencing platform supplies a sequencing service for plasmids or PCR products by Sanger sequencing using a machine (3730 DNA Analyzer, Applied Biosystem).
This platform sequences genes associated with different inherited disorders with a main focus on retinal disease: coding exons and flanking intronic regions of different genes are amplified with the use of polymerase chain reaction (PCR), purified by exo/SAP (exonuclease I/shrimp alkaline phosphatase), and sequenced with the use of a sequencing mix (BigDye Terminator v1.1 Cycle Sequencing Kit, Applied Biosystems) and analyzed on an automated 48-capillary sequencer (ABI 3730 Genetic Analyzer, Applied Biosystems).
The results are interpreted by a software application (SeqScape, Applied Biosystems).
The sequencing platform also validates variants identified by targeted next generation sequencing, whole exome and whole genome sequencing and performs cosegregation and control analysis for the respective variants.
In addition the service sequences plasmids of different researchers of the Institut de la Vision to validate the sequence after cloning or for cellular over-expression analysis.
Publications
- Identification of a Novel Homozygous Nonsense Mutation Confirms the Implication of GNAT1 in Rod-Cone Dystrophy.
Méjécase C, Laurent-Coriat C, Mayer C, Poch O, Mohand-Saïd S, Prévot C, Antonio A, Boyard F, Condroyer C, Michiels C, Blanchard S, Letexier M, Saraiva JP, Sahel JA, Audo I, Zeitz C. PLoS One. 2016 Dec 15;11(12):e0168271. doi: 10.1371/journal.pone.0168271. eCollection 2016. - Next-generation sequencing confirms the implication of SLC24A1 in autosomal-recessive congenital stationary night blindness.
Neuillé M, Malaichamy S, Vadalà M, Michiels C, Condroyer C, Sachidanandam R, Srilekha S, Arokiasamy T, Letexier M, Démontant V, Sahel JA, Sen P, Audo I, Soumittra N, Zeitz C.
Clin Genet. 2016 Jun;89(6):690-9. doi: 10.1111/cge.12746. Epub 2016 Mar 4. - Whole-exome sequencing identifies KIZ as a ciliary gene associated with autosomal-recessive rod-cone dystrophy.
El Shamieh S, Neuillé M, Terray A, Orhan E, Condroyer C, Démontant V, Michiels C, Antonio A, Boyard F, Lancelot ME, Letexier M, Saraiva JP, Léveillard T, Mohand-Saïd S, Goureau O, Sahel JA, Zeitz C, Audo I.
Am J Hum Genet. 2014 Apr 3;94(4):625-33. doi: 10.1016/j.ajhg.2014.03.005. Epub 2014 Mar 27.
Team
Christina Zeitz
Scientific supervisor
Email: christina.zeitz@inserm.fr
Christel Condroyer
Technical manager
Email: christel.condroyer@inserm.fr