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Team:British Columbia/NotebookMisc
From 2012.igem.org
To teammates: This will act as a log for miscellaneous lab work (e.g., competent cell, media, and plate production).
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May 28
Started making first batch of DH5α and EPI300 competent cells. See Competent Cell Production.
May 30
Continued production of competent cells. OD600 values were monitored as cells grew until when it reached 0.420 for DH5α and 0.443 for EPI300, upon which they were iced immediately.
May 31
Finished production of the competent cell protocol that was initiated on May 28. Competent cells were not flash-frozen with liquid nitrogen, but were placed directly into the -80ºC freezer. Samples of each cells were transformed then plated for verification.
June 1
Competent cells worked. Plates of competent cells grew on antibiotic plates.
Made 2 bottles of 400 mL M9 + glucose media.
June 5
Made EPI 300, BL21, and DH5α competent cells with plasmid pIJ790 in them.
-Ruichen
June 6
Learned how to electrophorese cells with biobricks (1-7B, 2-17F, 2-14N, 2-9B, 3-13M, 3-14E, 4-3I, 5-1A, 5-12O), and plated them on plates to grow colonies.
Tested kanamycin (Kan) plates with control.
-Ruichen
June 7
Retreated the plates previously spread, KAN plates worked and most of them grew into colonies or covered plates completely. (advised to spread the plates with 50µl recovered cells instead 100µl in the future for better colony identification).
Made more 10µg/ml ampicillin (Amp) and 34µg/ml chloramphenicol (Chlor), and more agar plates of Amp and Chlor (half bagful respectively).
Electrophoresed 1-1N, 2-2K, 1-5P, 2-21O using DH5α, and plated them.
-Ruichen
In the afternoon, we (Ruichen, Mehul, Grace) made many more K12 competent cells, which we put into storage. (The protocol is now on the wiki.) There was also talk from Jacob and John about buying $0.50 worth of gasoline/diesel from the nearest gas station to test solvent resistance of some of our cells. I left before that happened, though. (Would have been fun to see the attendant's reaction!)
- grace
June 20
Cam made competent EPI300 pIJ790 cells that were grown in excess arabinose. These were stored in the -80C freezer and have "Ep" labeled on the tube caps.
