Lab July 30 2009

From 2009.igem.org

10:00 am Derek

In the lab to start broth cultures of yesterday's rehydrated parts. It looks like there are good colonies for 3 out of the 4 parts. For some reason, neither plate of R0051, the lambda cI regulated promoter, showed any growth at all.

12:00 pm Layne

We nanospec'd the lambda cI regulated promoter to verify a good rehydration.

nanospec'd as 73.7ng/μL (2μL sample had, Abs = 1.197, A260 = 1.475, A280 = 1.537)

This seems to be in the same range as the RFP generator.

Eli used some solution from Dr. Nano's lab to blank the nanospec, we didn't have access to this today so we used water as the DNA was rehydrated in water.

The broth culture for the RFP part we rehydrated first looks good. I was at first put off by the very strange pinkish colour until I realized that of course that was the RFP.

3:00 pm Kyliah

We plated all of the remaining cells from yesterday's transformation attempt for part R0051 (pλcI) and put that in the incubator to grow up. If very few to no colonies form, then perhaps the plasmid rehydrant was not fully mixed. The higher reported concentration today might reflect that, if yesterday we pipetted mostly water.

The RFP generator was also subcultured earlier, so we could get a log-phase broth culture to freeze at -80oC: Cryopreservation Protocol.

We stored duplicate eppendorf tubes containing 20% glycerol for each of parts I13521 (RFP generator), J06504 (mCherry CDS), B0015 (double transcription stop) and B0034 (standard RBS).

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