Targeted Sequencing

Even with reductions in the cost-per-base of DNA sequencing, routine sequencing of large numbers of whole eukaryotic genomes is not always economically feasible. Enrichment of specific genomic regions, prior to sequencing, is an effective method for reducing sequencing costs and simplifying analysis. Targeted sequencing of large cohorts of samples is a powerful tool for the discovery and detection of disease-causing variants associated with many inherited diseases and cancers.

Hybrid capture and PCR-based methods are currently the most common methods for target-enrichment. Hybrid capture involves pools of oligonucleotide probes designed to target specific regions of interest within a fragment library. Non-specific hybrids are removed, resulting in a library enriched for the targeted DNA. Highly multiplexed PCR can also be used to generate amplicon pools for targeted sequencing. Each method has strengths and weaknesses, and the optimal strategy is often determined by the factors such as specificity, sensitivity, sample type and input, throughput, target size, turnaround time, sequencing platform and cost.

The construction of libraries with maximum molecular complexity and minimal bias is critical for targeted sequencing applications. KAPA library preparation and amplification kits utilize optimally-formulated and evolved enzymes, including KAPA HiFi DNA Polymerase, and optimized protocols to achieve superior library yields and limit amplification bias. KAPA Library Preparation and KAPA Hyper Prep Kits are compatible with the Roche® NimbleGen™ SeqCap EZ System, SureSelect Target Enrichment systems from Agilent Technologies, and xGen® Lockdown® probes from IDT.