A simplified hybrid capture approach retains high specificity and enables PCR-free workflow

We present a targeted sequencing workflow that eliminates bead-based capture, multiple washes, and post-hybridization PCR, while improving various aspects of data quality. The performance of the approach was evaluated by sequencing hundreds of samples and demonstrating high on-target rates, reduced duplicates, and improved indel accuracy. By combining the approach with a PCR-free library preparation, we enable an entirely PCR-free targeted sequencing assay which further improves indel calling, and shows the ability to call an HTT expansion, associated with Huntington's disease. This streamlined approach addresses key operational challenges in targeted sequencing, offering potential benefits for applications requiring rapid turnaround times or increased capability in variant detection.