Chronological Lab Notebook

Doesn't look like the electroporation worked. I did get one bit of contam tho which is nice... (Maybe one colony on one plate is legit? Or maybe they need more time? But I suspect I need to re-try with more rigorous cell prep :/

I got excited when I noticed that the flowers which I had injected with the Odin's agrobacterium Ruby system had some bright fluorescence under 360 nanometer UV light around the injection sites. There was a little bit of red ruby expression too, as I expected, but nowhere had I seen anything about a fluorescent reporter. This was exciting because I had used some DNA that I had extracted via miniprep from the agrobacterium in a first YOLO test of my Gene gun. I rushed to fish the test onion slices out of the dustbin and put them under the scope with a UV light illuminating them. Sure enough on the positive sample that I had shot, I could see some vague blue glow in a few cells similar to the pictures I saw online for the original O'Reilly Jean gun instructions. Very exciting! Except that I didn't know why they should be fluorescent genes in this plasma. I asked the Odin people and they confirmed that it shouldn't have any fluorescence- it's just Ruby. I am now tasting on a fresh flower, just the injection media or just water to see if the fluorescence is a response to damage by the primrose plant- my suspicion is that the onion was also some sort of autofluorescence and unrelated to the DNA that we shot into it. Update: yup, just the injection media was enough to cause some bright fluorescence in the petals - false alarm.

I'm testing out a bunch of different combinations of parameters for electroporation both in E coli and in the pseudomonas species that I have. Trying variants with a 1.2 mm Gap and a 2.4 mm Gap. Trying room temperature versus chilled trying one spark versus three and doing it all in a sort of taguchi array.

I got a boost converter than can go up to 60V. Poured a 1% agar gel in a clear plastic tub, with carbon rods as electrodes. Ran some food coloring and some highlighter fluid to test. 0.2% sodium bicarbonate buffer. 40V. One binder clip touched the buffer and gave some yucky reactions. But the dyes did separate. Proper agarose, TBE and seegreen safe DNA dye arrive in a few days.

Until I retroactively add these experiments manually

Isolating a glowy pseudomonas, trying to lyse cells, pouring a few plates

• First gram stain
• Had various things growing