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Team:Valencia/Project/Lab work/2 experiments
From 2008.igem.org
(Difference between revisions)
| Line 9: | Line 9: | ||
<h3> Materials </h3> | <h3> Materials </h3> | ||
| - | In our experiments we worked with the the following ''Saccharomyces cerevisiae'' strains | + | In our experiments we worked with the the following ''Saccharomyces cerevisiae'' strains kindly handed by [http://www.cbm.uam.es/mitolab/fichapersonal.aspx?idpersona=6 Eduardo Rial] : |
*UCP+ | *UCP+ | ||
*UCP- <!--control--> | *UCP- <!--control--> | ||
| Line 16: | Line 16: | ||
<h3> Method </h3> | <h3> Method </h3> | ||
| + | Each of our experiments entails the following steps: | ||
| + | <ol><li> We prepare a 30ml inoculum for each strains in 100ml Erlenmeyer flasks with [[Team:Valencia/Notebook/Protocols#SD Medium | SD medium]]. We leave it overnight in the 28ºC stove. | ||
| + | </li><li> We prepare a second inoculum for each strain, this time in with 100ml [[Team:Valencia/Notebook/Protocols#SP liquid medium | SP medium]] in 1L Erlenmeyer flasks. The initial O.D. of this second inoculum should be aproximately 0.2. | ||
| + | |||
| + | </li><li> We start the experiment in the LCCs with a volume of 100ml, [[Team:Valencia/Notebook/Protocols#SP liquid medium | SP medium]], and a initial O.D. of 0.6. | ||
| + | |||
| + | </li><li> We add 1% galactose to each of the LCCs containing the four yeast strains. This sugar is the UCP gene Gal7 promoter inductor. | ||
| + | |||
| + | </li><li> We monitor the temperature evolution in the LCCs thanks to our [[Team:Valencia/Project/Lab_work | LCC system]]. | ||
| + | |||
| + | </li></ol> | ||
Revision as of 11:55, 9 October 2008
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.
