Team:UCopenhagen/Notebook/Week 37
Week 37 (9th-15th of September)
12th September 2019
1. Setting-up the culture for the Bioactivity test the next day
Team members: Noël
100 µL of the positive culture that was created on the 29th August (S4C3 Yeast GPER-Galphas-STE12-ZsGreen) was put into fresh Glu-U media and left over night at 30° C. The colony was left in the fridge until its use.
16th September 2019
1. First Estradiol bioactivity assay
Team members: Noël and Hitesh
Materials
Procedure
- Dilute the cultures with the corresponding media (GPER-yeast->Glu-U, control-yeast -> YPD) to an OD600 of 0.5.
- Make Estradiol dilution
NOTE: A systematic error was made here, as the concentrations that were used here were way higher than the physiologically relevant ones!
The following solutions were mixed:1x stock: 1 M using EtOH as solvent
1000x diluted working solution: 1 mM (1µl of 1x stock + 999 µl EtOH)
- Filling the 96 well plate.
The following scheme samples were mixed:
Abbreviation Used Estrogen solution Volume Estrogen solution [µl] Volume water [µl] Used yeast culture Volume of yeast culture [µl] Total volume/well [µl] T0 - - 20 Transformed 120 140 T4 1000x 0.6 19.5 Transformed 120 140 T6 1000x 0.8 19.2 Transformed 120 140 T8 1000x 1.1 18.9 Transformed 120 140 T10 1000x 1.4 18.6 Transformed 120 140 T12 1000x 1.6 18.4 Transformed 120 140 T14 1000x 1.96 18.0 Transformed 120 140 T16 1000x 2.24 17.8 Transformed 120 140 T50 1000x 7 13 Transformed 120 140 T100 1000x 14 6 Transformed 120 140 T500 1x 0.7 19.3 Transformed 120 140 T1000 1x 1.4 18.6 Transformed 120 140 UN0 - - 20 Untransformed 120 140 UN4 1000x 0.6 19.5 Untransformed 120 140 UN6 1000x 0.8 19.2 Untransformed 120 140 UN8 1000x 1.1 18.9 Untransformed 120 140 UN10 1000x 1.4 18.6 Untransformed 120 140 UN12 1000x 1.6 18.4 Untransformed 120 140 UN14 1000x 1.96 18.0 Untransformed 120 140 UN16 1000x 2.24 17.8 Untransformed 120 140 UN50 1000x 7 13 Untransformed 120 140 UN100 1000x 14 6 Untransformed 120 140 UN500 1x 0.7 19.3 Untransformed 120 140 UN1000 1x 1.4 18.6 Untransformed 120 140 - For each of these samples triplets were loaded.
- The plate was left overnight at 30° C and shaking at 150 rpm.
Results
Because of a systematic error, the fluorescence in each well was to high and it was impossible to analyse the data properly.
About Us
We are Ovulaid: a team of 13 students from the University of Copenhagen working on a novel ovulation detection system, using synthetic biology.
Keep in Touch
iGEM Team Copenhagen
iGEM_Copenhagen
iGEM_Copenhagen
UCPH.IGEM2019@gmail.com
Address
University of Copenhagen
Thorvaldsensvej 40, Frederiksberg C
Denmark
