SureSelect DNA Capture Array
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Agilent’s SureSelect DNA Capture Array provides an alternate approach to target enrichment that is suitable for smaller studies. Similar to the SureSelect Target Enrichment System, the DNA Capture Array is also capable of isolating user-defined genomic regions from complex genomes, significantly lowering sequencing costs by focusing only on regions of interest. This array-based target enrichment method is ideal, however, for researchers who need to test a small number of samples with various capture designs.
Figure 1. SureSelect DNA Capture Array assay followed by DNA sequencing.
The above figure illustrates the SureSelect DNA Capture Array method. In a typical capture, targeted regions (yellow) are tiled by 60-mer probes (green). Fragmented DNA is ligated to adapters (red), size-selected, and amplified. DNA is hybridized, washed in stringent buffer, and the bound fraction eluted by heat treatment. Eluted DNA can be further amplified, loaded onto an Illumina Genome Analyzer and sequenced. The SureSelect DNA Capture Array is designed to achieve optimal results with the Illumina Genome Analyzer system, but can be used with any sequencing system with simple modifications to the protocol.
In a collaborative effort, Agilent used 244K SureSelect DNA Capture arrays to capture exonic regions relevant to a breast cancer sequencing study. Roughly ~0.025% of the human genome was captured using 60-mer probes, including 1,287 discrete genomic regions, then released and sequenced. Ultimately, a 2700-fold enrichment of sequencing reads within targeted regions was obtained. The results validated the SureSelect DNA Capture Array application as effective. Sequencing reads covered over 99.8% of the targeted regions and 98% of the targeted bases with at least one read. The normalized average per-base read depth was 27 per million 32-base reads. These results confirm that Agilent’s SureSelect DNA Capture Arrays can provide a rapid and effective solution for targeted sequencing of genomic regions of interest, particularly for smaller studies.
Human All Exon Kit now available.
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