Retrogen Custom SNP Discovery & Resequencing Service
Single nucleotide polymorphisms (SNPs) are the most common variation responsible for genetic diversity between individuals, accounting for more than 85% of the variability. Recent advances in SNP identification and analysis have made SNP genotyping an invaluable tool to examine the variations responsible for disease susceptibility and drug responsiveness. SNP genotyping applications have recently extended into personalized health care, diagnostics, forensics, animal studies and agriculture. In order to address the needs of this rapidly growing SNP market, we at Retrogen employ a suite of integrated solutions to assist the customer accomplishing their SNP discovery projects from beginning to the end.
Our SNP Discovery and Resequencing service starts out with the assay development by using commercial and proprietary informatics to align the cDNA sequence to the genomic sequence and choose a specific region for designing the primers. The primers are selected across the entire span of the targeted exon and are synthesized in our oligo synthesis department. All primer design and synthesis have to go through rigorous quality control to ensure that all the amplicons have sequencing data greater than Phred20 score. Exons are amplified from genomic DNA, and the resulting amplicons are purified for sequencing. The sequencing data is collected using fluorescent dye-terminator chemistry by fully automated ABI 3730xl sequencers. The electrophereograms are assembled with reference sequence and the potential SNPs are scored. Candidate SNPs with Phred20 score are validated by re-sequencing the opposite DNA strand to confirm the polymorphism.
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