Today was not much of a departure from yesterday. After we left our slides hybridize overnight, we began this morning by washing them in a series of SSC (saline sodium citrate). We then incubated the slides in blocking buffer, which prevents the antibody from targeting anything else other than our probe. Next, we incubated the slides with our antibody that targets the DIG on our probes. After letting them go for two hours, we added nitro blue tetrazolium and 5-Bromo-4-chloro-3-indolyl phosphate (or BCIP). These are catalyzed by the phosphotase on the antibody to form a blue dye, which allows us to see where our probes hybridized. Unfortunately, we didn’t see any development yet but we’re leaving it overnight to see if it works!
We also got our game plan down for tackling the sea star transcriptome! We’re aiming to investigate different enriched processes and the differentially expressed genes in them to see whats up with the sick stars!