Team:Valencia/Notebook/July

From 2008.igem.org

July 1st

- We prepared some glycerol stock from the four Saccharomyces cerevisiae strains we received from [http://www.cbm.uam.es/mitolab/fichapersonal.aspx?idpersona=6 Eduardo Rial] :
      Negative control, UCP -.
      Strain containing UCP1, UCP +.
      Mutant stains over expressing UCP1, Gly 76Δ and Gly 175Δ.

- We calibrated our calorimeters with hot water.

July 2nd

- We prepared our first SD medium.

- We calibrated our calorimeters with hot water.

July 4th

- We prepared some plates for inoculum. They were stored a the 28ºC stove.

July 7th

- We prepared some inoculum in Erlenmeyer flasks for our experiment the following day. We kept them overnight in a 30ºC shaking stove.

July 8th

- The experiment consisted on:
SD medium
Four strains
We left it overnight, but the air conditioning went off and the room temperature rose.

July 9th

- We characterized culture growth in our calorimeters by measuring DO, with just one strain.

July 10th

- Trying to see if there is basal rate of UCP1. Experiment:
SD medium
Calorimeters containing: water, medium, UCP-, Gly 175Δ.

July 11th

- It looks like our Saccharomyces cerevisiae mutant strain has a basal rate of UCP1... We repeated the experiment to be sure.

July 12th

- We obtained the same results.

July 14th

- The plate is completely dry,the water evaporeted...WHY???? the first unsolved mystery of valencia iGEM 2008.

July 15th

- We prepared inoculum in Erlenmeyer flasks. We kept them overnight in a 30ºC shaking stove.

July 16th

- Trying to find out which is the best moment for activating UCP1.... The glucose in the medium must be consumed by the induction time. DO at the beginning of the experiment was 0.05.
Experiment:
       SP medium
       Galactose activation.
       One strain.
       Activation at two, three and four hours after inoculating. D.O. was measured at each time.

July 17th

- Our lab electricity went off during the night. How many experiments will have it ruined?

July 18th

- We prepared lots of plates with our SD medium...Unfortunately, we forgot to add glucose.

- We carried out the pilot experiment:
      SP medium.
      Galactose activation at 3 hours (approximate DO 0.15).
      Four strains.
We see weird oscillations.

July 20th

- We stopped the shaking and carried out the experiment again to see whether the oscillations were related to the shaking.

July 21st

- We carried out the pilot experiment from July 18th again. Weird results, we need to repeat it.

July 22nd

- We repaired our calorimeters. They were the reason for yesterday weird results.

July 23rd

- We repeated the pilot experiment from July 18th.

July 24th

- Our male group members decided to carried out their own particular experiment involving parafilm, plasticine and teeth filling paste... They wanted to find out if the oscillations would stop if they covered the thermocouples with different materials they found in the contiguuos labs, or immerse them in the liquid.
Meri, who is in charged of the lab, wasn't very happy the following day...

July 25th

- Another curious experiment was carried out: one thermocouple was covered with plasticine while the others weren't.

July 26th

- One of our calorimeters wasn't working properly.

July 28th

- We repaired the calorimeter and calibrated once again.

July 29th

- Our cultures haven't grown enough for the experiment. Temperature and medium were all right, we'll have to wait...