In situ HCR in non-traditional arthropods

Visualizing the expression of genes is a fundamental tool in molecular biology. Traditional colorimetric in situ hybridization using long RNA probes has been a staple for visualizing gene expression but has many drawbacks. In situ HCR v3.0, developed by Choi et. al. 2018, offers improvements over traditional in situs in nearly every aspect: probes can simply be ordered rather than painstakingly cloned and transcribed, which also makes them cost-effective; an HCR takes just three days to complete rather than five or more days; HCR is robust and works well for first-time users; and HCR probes can be multiplexed, allowing four to eight genes to be visualized in a single sample. HCR has been used successfully in many arthropods, including insects (Drosophila, Tribolium), crustaceans (Parhyale, Daphnia, Artemia), and chelicerates (Limulus horseshoe crab, Acanthoscurria tarantula). In this demo, you will learn how to design and order HCR probes as well as best practices for experimental design.