From OpenWetWare
Jump to navigationJump to search


It's been a long time since I've been in lab, though that should change in the upcoming weeks. I'll try to finish about 5 westerns by the end of the semester, and depending on the amount of time that I have, I may also work into reading period (after my 91r is due).

Next up: plans for a second experimental Western blot.

Key differences:

  • Inclusion of a different primary antibody that detects levels of a protein that is consistently produced in E. coli
    • This antibody is limited to the following bio-tech companies:
      • Invitrogen
      • Sigma
      • Santa Cruz
    • A good place to start is here
  • Instead of taking time points every half-hour, instead we are going to be taking time points once every 3 hours.
    • Justification:
      1. It will be less time consuming to collect data points over the span of 1 day
      2. The data will fit on the gel much easier (allowing for two positive controls: J36834 (KaiC + promoter) and J36335 (KaiA, KaiC + promoter)
  • In the end, I'll be using s70 as the protein that will be detected. The shipment for the protein should arrive sometime next week.

What's next: Planning a second experiment for a collection of data, probably including an interaction part and an oscillation portion. This will be done after the Western Blot with the primary antibody is completed (around April 3rd). The experiment will probably proceed from April 4-5th, and the projected goal for completion of the Western for this project will be complete around April 7th.