Team:Valencia/Project/Lab work/2 experiments
From 2008.igem.org
2.-Demonstration that thermogenin-expressing yeast strains can heat their own broth medium.
Materials
In our experiments we worked with the the following Saccharomyces cerevisiae strains kindly handed by Eduardo Rial :
- UCP+
- UCP-
- Gly175Δ
- Gly76Δ
Method
Each of our experiments entails the following steps:
- We prepare a 30ml inoculum for each strains in 100ml Erlenmeyer flasks with SD medium. We leave it overnight in the 28ºC stove.
- We prepare a second inoculum for each strain, this time in with 100ml SP medium in 1L Erlenmeyer flasks. The initial O.D. of this second inoculum should be aproximately 0.2.
- We start the experiment in the LCCs with a volume of 100ml, SP medium, and a initial O.D. of 0.6.
- We add 1% galactose to each of the LCCs containing the four yeast strains. This sugar is the UCP gene Gal7 promoter inductor.
- We monitor the temperature evolution in the LCCs thanks to our LCC system.