Results
![](https://static.igem.org/mediawiki/2016/2/26/T--LambertGA--purpleline.jpg)
Inoculations of GFP Constructs (58 tubes)
After making our constructs, we inoculated cultures from previous transformations that have successfully expressed the fluorescent protein.
1.P-Lambda-R--LacI--GFP (no deg tag)
- 3 tubes in each cell type (DH10, Keio Wild, Keio ClpP) induced with 0 uM IPTG (plain LB)
- 3 tubes of each cell type (DH10, Keio Wild, Keio ClpP) induced with 100 uM IPTG (plain LB)
- RESULTS:
- 3 tubes in each cell type (DH10, Keio Wild, Keio ClpP) induced with 0 uM IPTG (plain LB)
- 3 tubes of each cell type (DH10, Keio Wild, Keio ClpP) induced with 100 uM IPTG (plain LB)
- RESULTS:
- 3 tubes in each cell type (DH10, Keio Wild, Keio ClpP) induced with 0 uM IPTG (plain LB)
- 3 tubes of each cell type (DH10, Keio Wild, Keio ClpP) induced with 100 uM IPTG (plain LB)
- RESULTS:
5. RESULTS:
- 10/17: The GFP constructs were brought to the plate reader at Georgia Tech. Although cells were grown in the liquid culture, they did not fluoresce.
![](https://static.igem.org/mediawiki/2016/e/e8/T--LambertGA--platereaderresults.jpg)
![](https://static.igem.org/mediawiki/2016/c/c5/T--LambertGA--inoculationtubes.png)
![](https://static.igem.org/mediawiki/2016/8/8f/T--LambertGA--dyanmicduo.png)
Troubleshooting for Lack of Proper Tube Labeling
We plated our constructs in all three cell types(DH10, Keio Wild, and Keio ClpP) on Kanamycin, Tetracycline, Chloramphenicol, and Ampicillin. We were testing to verify what backbones the plasmids were in. As the image shows, our cells grew in most of the plates resistant to the specific antibiotics. We are in the process of figuring out how we obtained these results, but we hypothesized that our cells contain constructs with all the vectors with backbones resistant to those antibiotics.
![](https://static.igem.org/mediawiki/2016/c/c7/T--LambertGA--platestotest.png)
Sequencing Results
![](https://static.igem.org/mediawiki/2016/d/de/T--LambertGA--sequence1.jpg)
The R0011 ClpX part of our DNA construct was sent in for sequencing and confirmed to be matching DNA through Eurofins MWG Operon.
![](https://static.igem.org/mediawiki/2016/c/c2/T--LambertGA--sequence2.jpg)
The ClpP B0033 CI part of our DNA construct was also sent in for sequencing and confirmed to be matching DNA through Eufofins MWG Operon.
Expected Results
Medals
![](https://static.igem.org/mediawiki/2016/1/11/T--LambertGA--bronzemedal.png)
![](https://static.igem.org/mediawiki/2016/5/55/T--LambertGA--silvermedal.png)
![](https://static.igem.org/mediawiki/2016/d/dc/T--LambertGA--goldmedal.png)
![](https://static.igem.org/mediawiki/2016/b/b6/T--LambertGA--checkbox.jpg)
![](https://static.igem.org/mediawiki/2016/b/b6/T--LambertGA--checkbox.jpg)
![](https://static.igem.org/mediawiki/2016/b/b6/T--LambertGA--checkbox.jpg)
![](https://static.igem.org/mediawiki/2016/b/b6/T--LambertGA--checkbox.jpg)
![](https://static.igem.org/mediawiki/2016/b/b6/T--LambertGA--checkbox.jpg)
![](https://static.igem.org/mediawiki/2016/b/b6/T--LambertGA--checkbox.jpg)
![](https://static.igem.org/mediawiki/2016/b/b6/T--LambertGA--checkbox.jpg)
Silver Gold