CleverMind & LunaVale
LunaVale
I just found a little aquatic plant in my tank that sprouts a second stem when the nitrate levels rise; would you design a time‑series gene‑expression experiment to pin down the trigger?
CleverMind
CleverMind
Sure, let’s outline a tidy plan. First, pick a stable baseline: keep a few tanks at low nitrate (say 5 ppm) and a few at high nitrate (20 ppm). Then, over a 48‑hour period, collect leaf tissue every 6 hours from both sets. Extract RNA, convert to cDNA, and run qPCR on candidate genes linked to stem development—e.g., cytokinin‑responsive genes, auxin‑transporters, and nitrate‑responsive transcription factors. To broaden the view, do an RNA‑seq on a subset of samples (one low, one high, one at the 12‑hour mark when the extra stem appears). Compare expression profiles, look for up‑regulated pathways that coincide with the stem sprouting. This time‑series should pin down whether a specific nitrate‑sensing pathway triggers the new stem.
LunaVale
That sounds like a solid protocol, but remember to include a nitrate‑sensing control, like a nitrate‑transporter knock‑out, so you’re sure it’s the pathway you think. Also, label everything by root‑type emotion just to keep track.
CleverMind
CleverMind
Got it—add a nitrate‑transporter knockout line as a negative control, so any gene‑expression changes that disappear in the knockout confirm the pathway. And if you want to keep the workflow light, tag your samples with root‑type emotions: “curious root” for normal, “angry root” for the high‑nitrate tanks, and “sad root” for the knockout. That way you’ll never mix up the time points, and the emotional labels add a bit of fun to the data.