Illumina Target Enrichment Workflow

Learn the fundamentals of Next Generation Sequencing (NGS) and compare two targeted sequencing methods: amplicon-based and enrichment-based approaches. Enrichment sequencing is highlighted for its ability to handle larger gene panels and comprehensive variant profiling.

Use bead-linked transposomes (BLT) to simultaneously fragment the DNA sample, normalize it, and ligate sequencing adapters. This process provides uniform coverage with minimal region-specific bias.

Anneal index adapters to the DNA through reduced cycle amplification at the adapter regions created during tagmentation.

Combine 1 to 12 multiplexed samples and denature them in preparation for hybridization with targeted probes.

Add biotin-labeled probes specific to target regions and streptavidin-coated beads to bind and isolate the targeted DNA fragments from the pooled sample.

Magnetically pull down the enriched DNA fragments and elute them from the streptavidin-coated beads. The complete library preparation workflow takes 6.5 hours and is automation-compatible.

Run validated and quantified libraries on any Illumina sequencing platform, adjusting multiplexing based on desired coverage depth and platform output capacity.

Process sequencing data using Dragon enrichment application on NextSeq 1000/2000, on-premise servers, or cloud platforms to perform secondary analysis including SNV/indel calling, copy number variation, and structural variant detection.