5/18

Starter Culture from 5/17 had an OD 600 of 3.1 for tube #1 and 3.0 for tube #2
- This is too high, as the desired range is around 0.5 to 1.5.
Remaking starter culture. Picked 3 colonies from plate #2 (P+S 2) and incubated in 3 tubes with 10 ml LB and 10 ul 1000X chloramphenicol
- Incubated at 37C starting at noon. (200 rpm)
- At 2 pm, the OD was still essentially 0.
- So apparently, I was reading the spec incorrectly. I was using cuvettes, but I had forgotten to change the setting to "10 mm cuvettes"
- At 7:30 pm, (7.5 hours of incubation), the OD600 was 1.21
  - This is a little too high, as 0.8 is about optimal, but I went with it anyways.
Added 5 ml of the starter culture to 245 ml Difco LB and 250 ul of 1000X chloromphenicol to a 1L flask with the little ridges in the bottom for better aeration (baffled flask)
Incubate at 37C 200 rpm starting at 8 pm
- At 10:40pm OD600 = 0.336
- At 11:05pm OD600 = 0.441
At this point I added 1 ml of 100 mM IPTG to make 0.4 mM final IPTG concentration.
Placed back into the incubator starting at 11:10 PM

When Megan did the miniprep from the pET 24a transformed bacteria, the yield was very low.
I re-picked 2 colonies from the pET 24a transformed plate I made on 5/10 and inoculated them on 5/17.
- One tube didnt seem to have any growth and the other tube had very precipitated bacteria.
- The yield of the miniprep from tube #2 was very low.

That plate from 5/10 seems funky, so Megan will re do the transformation, with less DNA and will make a 1:10 dilution plate as well

Team:UCLA/Notebook/Honeybee Silk/18 May 2015