It had been two days since the transformation yesterday, and I had not seen any growth on my plates. I was becoming concerned. It can be tricky to orchestrate the teaching and the actual transformation procedure, and I have had less than optimal results with this media, strain, and plasmid before. I was not sure what I would do on Sunday if there were no transformants! It would be a disaster, and very demoralizing.
I had a meeting to go to that would result on me being on the red line, so on the way back I stopped by and took a look at the cultures with my blue LED. Nothing was growing, except for what appeared to be some e. coli growing on the ampicillin plate, which I took to be an ill omen. I left my blue LED and some batteries there, because I figured that if there were transformants, they would be more likely to grow on the multitude of plates at sprout instead of the two that I had.
I wasn’t sure what to do there, so I grabbed a few items for doing another transformation and headed home, convinced that I would have to do another transformation. Upon my return home, I obsessively checked again.
When I returned from dinner, I decided to take one last look. I was greeted with two plates full of transformants, in perfect little green colonies. Having left my sole blue LED at sprout, I initially thought that I would have no way of testing the fluorescence, until I spied my arduino nano on my desk. I remembered that it had a blue LED on it, so I plugged it into my laptop and used it to light the plates, which indeed fluoresced.
Lessons learned: Be patient. Synch your life to the organism you are studying, not the other way around. Also arduinos are good for many things.
The final question you may have is “why did they take so long to grow?”. The answer there is something I should have recognized! I totally forgot that there is often a “lag phase” of growth in bacteria. This is the phase where bacteria are generating the needed metabolites and substrates to adjust to their surroundings. The colonies are also growing (in this case) from a single bacteria! So it makes sense that there was a pretty big lag from when they were plated, until I could see them.