From SELEX to Sequencing: Uncovering enriched Aptamers with Next-Generation Sequencing and Bioinformatics.

An optimized NGS-guided SELEX computational workflow identifies target-specific aptamer candidates by clustering enriched sequences, yielding six clusters after removing negative-pool sequences and seven positive clusters.

• Why it matters: SELEX can enrich aptamers, but prioritizing true target binders among nonspecific sequences is challenging; integrating NGS aims to improve identification and selection.
• What they did: The workflow includes sequencing library preparation, preprocessing, clustering, multiple sequence alignment, phylogenetic analysis, and secondary structure prediction, using two comparative strategies across positive and negative selection pools.
• The result: Candidates prioritized by secondary structure, sequence abundance, and genetic distance from nonspecific binders showed strong specific binding to the target protein in dot blot assays, supporting streamlined downstream validation.