Sequence Capture
NimbleGen Sequence Capture technology is a revolutionary process for the enrichment of selected genomic regions from full complexity human genomic DNA in a single step. Sequence Capture was developed to eliminate the necessity of setting up thousands of PCR reactions, instead allowing for parallel enrichment of target regions in a single experiment.
Roche NimbleGen created, further refined, and optimized large-scale genomic enrichment technology. Our company is at the forefront of targeted sequencing, driving the innovation of Sequence Capture technologies including custom capture. Capitalizing on the efficiencies inherent with parallel enrichment, researchers can now design economical, higher throughput, and time-saving next-generation sequencing experiments. In combination with high throughput sequencing (short- or long-read), Sequence Capture has made targeted sequencing possible and accessible to more life science researchers. We currently offer the following option for targeted enrichment of genomic regions:
- SeqCap EZ Library is a solution-based capture method that enables enrichment of the whole exome or customer regions of interest in a single test tube. An automated solution for SeqCap EZ Library using the Caliper Sciclone NGS workstation is now available. With the Sciclone NGS workstation you can process up to 288 samples per week.
Protocol
Roche NimbleGen offers two types of capture methods: SeqCap EZ Library, a solution-based method and Sequence Capture Arrays, an array-based capture method.
Sequence Capture Protocols
- Genomic DNA: SeqCap EZ Oligo pool or an array is made against target regions in the genome.
- Library Preparation: Standard shot-gun sequencing library is made from genomic DNA.
- Hybridization: The sequencing library is hybridized to the SeqCap EZ Oligo pool or to the Sequence Capture array.
Steps 4 and 5 are different for each protocol:
SeqCap EZ Library, biotinylated DNA oligos in solution
- Bead Capture: Streptavidin beads are used to pull down the complex of capture oligos and genomic DNA fragments.
- Washing: Unbound fragments are removed by washing.
Sequence Capture, capture probes synthesized on array:
- Washing: Unbound fragments are removed by washing.
- Target Fragment Elution: The enriched fragment pool is eluted and recovered from the array.
- Amplification: Enriched fragment pool is amplified by PCR.
- Enrichment QC: The success of enrichment is measured by qPCR at control loci.
- Sequencing-Ready DNA: The end product is a sequencing library enriched for target regions, ready for high throughput sequencing.
