DNA seq of the polyT oligopool hybridized against a specific target
To enable high-throughput quantification of probe-target interactions under these conditions, we developed a novel and scalable hybridization and sequencing workflow. Biotinylated targets were immobilized on streptavidin-coated beads and hybridized with a probe pool under defined temperature and salt conditions. All probes contained Nextera adapter sequences to allow Illumina library preparation and sequencing. Following stringent washes, probes were eluted, amplified, sequenced, and quantified. Sequencing read counts provided a direct, quantitative measure of probe-target binding, enabling systematic evaluation of affinity and specificity across probes.