Single Nucleotide Polymorphisms (SNPs) are the most common type of sequence variation and account for about 90% of sequence differences in humans. Of particular interest are coding SNPs (cSNPs), which have the potential to alter the coding sequence of genes and possibly gene functionality. SeqWright offers customized SNP discovery packages for targeting and characterizing sequence variations in large numbers.

Our SNP discovery service is designed to give customers a complete beginning-to-end solution. Assay development is accomplished using commercial and proprietary informatics tools, which align cDNA sequence to genomic sequence and choose amplicons with corresponding primer sets. Primers are designed to yield Phred20 data across the entire span of targeted exon and are manufactured in our DNA synthesis laboratory. Exons are amplified from genomic DNA, and resulting PCR products are purified for downstream processing. Data is collected using fluorescent dye-terminator chemistry by fully automated ABI 3730xl sequencers. Traces are compiled into contigs (when applicable) and scored by two independent observers using Phred base call confidence scores to identify high probability heterozygotes. Candidate SNPs are validated by re-sequencing the opposite DNA strand to confirm the polymorphism. Mapping against public databases is also available. All work is performed according to GLP and reviewed by an independent quality assurance (QA) unit.

Customers may enter our SNP discovery pipeline at any point from assay construction to sequencing. Genotyping is available for polymorphism verification and heterozygosity determination. Upon completion of the project, customers will receive a hardcopy report detailing the project and identifying all mutations/heterozygotes present in the sequence.

Features of the service include: · Assay development and validation · Primer design and synthesis · PCR amplification and purification · Automated fluorescent dye-terminator sequencing · Sequence trace alignment and editing · In silico mutation/variation identification · Heterozygosity detection via genotyping (if requested) · Hard copy final report