Solution-based targeted genomic enrichment for precious DNA samples

• Main Authors: Shearer Aiden, Hildebrand Michael S, Smith Richard JH
• Format: Article
• Language: English
• Published: BMC 2012-05-01
• Series: BMC Biotechnology
• Subjects: Targeted genomic enrichment; Sequence capture; Massively parallel sequencing; Illumina
• Online Access: http://www.biomedcentral.com/1472-6750/12/20

<p>Abstract</p> <p>Background</p> <p>Solution-based targeted genomic enrichment (TGE) protocols permit selective sequencing of genomic regions of interest on a massively parallel scale. These protocols could be improved by: 1) modifying or eliminating time consuming steps; 2) increasing yield to reduce input DNA and excessive PCR cycling; and 3) enhancing reproducible.</p> <p>Results</p> <p>We developed a solution-based TGE method for downstream Illumina sequencing in a non-automated workflow, adding standard Illumina barcode indexes during the post-hybridization amplification to allow for sample pooling prior to sequencing. The method utilizes Agilent SureSelect baits, primers and hybridization reagents for the capture, off-the-shelf reagents for the library preparation steps, and adaptor oligonucleotides for Illumina paired-end sequencing purchased directly from an oligonucleotide manufacturing company.</p> <p>Conclusions</p> <p>This solution-based TGE method for Illumina sequencing is optimized for small- or medium-sized laboratories and addresses the weaknesses of standard protocols by reducing the amount of input DNA required, increasing capture yield, optimizing efficiency, and improving reproducibility.</p>