Oligo synthesis for NGS in and out of the clinical setting

Scott Rose, Ph.D.

Director Of Molecular Genetics at Integrated DNA Technologies

Oligo synthesis for NGS in and out of the clinical setting Targeted pull-down enrichment using oligonucleotide probes has become an important tool for those researchers interested in isolating a smaller subset of loci defining a particular pathway or disease state. Using hybridization probe-based enrichment has its advantages over PCR based methods. Probes that are long enough, approximately 120 nucleotides in length are impervious to smaller base variations which allows for discovery of undocumented SNPs. Unlike PCR amplification, deletions or rearrangements can be detected without any additional information about chromosomal location. The improved on target capture percentage through the use of blocking oligonucleotides as part of the hybridization step has been previously demonstrated. A new generation of modified blocking oligonucleotides shows 10-25% increase of "on target captures" using libraries made with TruSeq(tm) adapters. Furthermore these new blockers have universal barcoded domains alleviating the need for individual barcode specific blockers.