OS-Seq

OS-Seq

Oligonucleotide-selective sequencing (OS-Seq)  was developed to improve targeted resequencing, by capturing and sequencing gene targets directly on the flow cell. In this method, target sequences with adapters are used to modify the flow cell primers. Targets in the template are captured onto the flow cell with the modified primers. Further extension, denaturation, and hybridization provide sequence reads for target genes. Deep sequencing provides accurate representation of reads.

Pros:

• Can resequence multiple targets at a time
• No gel excision or narrow size purification required
• Very fast (single-day) protocol
• Samples can be multiplexed
• Reduced PCR bias due to removal of amplification steps
• Avoids loss of material

Cons:

• Primers may interact with similar target sequences, leading to sequence ambiguity

• OS-Seq: Schmitt M. W., Kennedy S. R., Salk J. J., Fox E. J., Hiatt J. B., et al. (2012) Detection of ultra-rare mutations by next-generation sequencing. Proc Natl Acad Sci U S A 109: 14508-14513

1. Markkula N., Suvisaari J., Saarni S. I., Pirkola S., Pena S., et al. (2015) Prevalence and correlates of major depressive disorder and dysthymia in an eleven-year follow-up--results from the Finnish Health 2011 Survey. J Affect Disord 173: 73-80
2. Vattulainen S., Aho J., Salmenperä P., Bruce S., Tallila J., et al. (2015) Accurate genetic diagnosis of Finnish pulmonary arterial hypertension patients using oligonucleotide-selective sequencing. Molecular Genetics & Genomic Medicine 3: 354-362
3. Hopmans E. S., Natsoulis G., Bell J. M., Grimes S. M., Sieh W., et al. (2014) A programmable method for massively parallel targeted sequencing. Nucleic Acids Res 42: e88