"
Team:Chalmers-Gothenburg/Results
From 2012.igem.org
m (→Deletion of CWP2 gene) |
m (→Survival of yeast in urine) |
||
| Line 68: | Line 68: | ||
=='''Survival of yeast in urine'''== | =='''Survival of yeast in urine'''== | ||
| - | + | [[File:Growth_curve_urine.jpg|thumb|250px|Figure 1: Growth curve for cells in urine and YPD medium. The OD remains the same in the urine i.e. the cells are not growing.]] | |
| - | An initial test was performed in order to test the survival of yeast cells in urine, which is an important property of our biosensor if it should function as a pregnancy test kit. Pre-cultures of the lab yeast strain CEN.PK 113-11C were prepared in 5 ml YPD and grown O/N at 30 °C. The day after, the cells were centrifuged, resuspended in 40 ml YPD and 40 ml filter sterilized urine respectively and grown in shake flasks at 30°C. | + | An initial test was performed in order to test the survival of yeast cells in urine, which is an important property of our biosensor if it should function as a pregnancy test kit. Pre-cultures of the lab yeast strain CEN.PK 113-11C were prepared in 5 ml YPD and grown O/N at 30 °C. The day after, the cells were centrifuged, resuspended in 40 ml YPD and 40 ml filter sterilized urine respectively and grown in shake flasks at 30°C. OD<SUB>600</SUB> measurements were taken at two different points of time during the day. The growth curve can be seen in Figure 1. The cells in urine did not grow; the OD remained the same during the whole day, while the cells in YPD were proliferating. |
[[File:Spottest-urine2.jpg|thumb|250px|Figure 2: Spot test analysis of cells grown in YPD and urine for 4h. The cells survive in the urine and grow normal after being plated on a YPD plate.]] | [[File:Spottest-urine2.jpg|thumb|250px|Figure 2: Spot test analysis of cells grown in YPD and urine for 4h. The cells survive in the urine and grow normal after being plated on a YPD plate.]] | ||
| - | In order to test if the cells remained alive in the urin, 300 µl of each culture (after 4h) were taken and | + | In order to test if the cells remained alive in the urin, 300 µl of each culture (after 4h) were taken and OD<SUB>600</SUB> was adjusted to 0.5. The samples were diluted 3x, 9x, 27x and 81x and 10 µl aliquots of each dilution were spotted on an YPD plate (Figure 2). The cells from both the YPD and the urine media could grow, this means that the cells survived 4 h in urine and grew normal after being spotted on a YPD plate. Summarizing, yeast cells are unable to proliferate in the urine medium but do survive under these conditions. |
=='''Deletion of ''CWP2'' gene'''== | =='''Deletion of ''CWP2'' gene'''== | ||
Revision as of 21:51, 24 September 2012
Contents |
Results Summary
Survival of yeast in urine
An initial test was performed in order to test the survival of yeast cells in urine, which is an important property of our biosensor if it should function as a pregnancy test kit. Pre-cultures of the lab yeast strain CEN.PK 113-11C were prepared in 5 ml YPD and grown O/N at 30 °C. The day after, the cells were centrifuged, resuspended in 40 ml YPD and 40 ml filter sterilized urine respectively and grown in shake flasks at 30°C. OD600 measurements were taken at two different points of time during the day. The growth curve can be seen in Figure 1. The cells in urine did not grow; the OD remained the same during the whole day, while the cells in YPD were proliferating.
In order to test if the cells remained alive in the urin, 300 µl of each culture (after 4h) were taken and OD600 was adjusted to 0.5. The samples were diluted 3x, 9x, 27x and 81x and 10 µl aliquots of each dilution were spotted on an YPD plate (Figure 2). The cells from both the YPD and the urine media could grow, this means that the cells survived 4 h in urine and grew normal after being spotted on a YPD plate. Summarizing, yeast cells are unable to proliferate in the urine medium but do survive under these conditions.
Deletion of CWP2 gene
Expression of human LH/CG receptor
Introduction of indigo synthesizing genes
