Send A Specimen
To request a test, the referring physician must download and complete Test Requisition Form, and send it with the Specimen. The instructions for Specimen Collection and Handling are provided: Specimen Collection and Handling
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Our Protocol (Outlines)
Targeted DNA enrichment of selected regions of interest
Purified genomic DNA will be used to generate enriched library of the exons and the flanking intronic regions (10 to 30 bp) of the genes of interest. The library will be generated using Sure Select Target enrichment system provided by Agilent Technologies.
Sequencing of the enriched libraries on Solid 4 platform
The enriched DNA libraries will be sequenced on SOLiD 4 platform. SOLiD 4 platform enables generation of 100GB high-quality sequence data per run, with high accuracy based on dinucleotide-produced signals (as opposed to single-nucleotide-based signals).
The resulting sequence datasets are subjected to primary and secondary analysis.
Primary analysis aims to assure proper data production and includes alignment of the sequencing data against short (up to 100 megabases) reference sequence. It is performed by manufacturer provided (Applied Biosystems) software, integrated in the Solid 4 computer system.
Secondary analysis includes fine reading and assembly of all the generated sequencing reads, variation analysis, and analysis of the pathogenic potential of the identified variants. For optimal performance, secondary analysis can be run on several platforms in parallel:
- Bioscope Integrated hardware-software package (Applied Biosystems)
- Bowtie: short read aligner optimized for ultrafast performance on human data.
- Additional analyses software may be use, as needed.
For diagnostic purposes, all the identified variations of interest will be confirmd with by-directional Sanger sequencing.
After completion of the analysis on the PI-requested genes and confirmation of the variations of interest, the sequence results on the inquired genes will be reported to the requesting PI under the CLIA protocol