Short tandem repeat (STR) DNA profiling, having applications in e.g. the fields of forensics and diagnostics, is currently done based on PCR amplification followed by amplicon sizing using capillary electrophoresis. This labor- and time-intensive methodology requires expensive lab equipment. The demand for forensic DNA profiling at the crime scene or at police stations is increasing. To tackle this issue, we recently devised a novel type of dequenching based DNA hybridization probes, and demonstrated proof-of-concept applicability in melting curve based STR profiling. In this project, we propose to perform a thorough characterization of probe hybridization, to gain an in-depth understanding of the hybridization process and hence improve the probes by rational design. We aim to understand the influence of the presence of PCR amplification artifacts on the discriminatory power of the probes for STR profiling. Furthermore, a more advanced hybridization probe concept will be explored. Ultimately, we will study the effect of probe attachment to a surface on hybridization efficiency, devising the best immobilization strategy to enable implementation in a microarray format. We plan to consolidate this knowledge by demonstrating on-chip application, using both singleplex and multiplex chip designs.