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Team:UT Dallas/immunobot results
From 2011.igem.org
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<h2>Results</h2> | <h2>Results</h2> | ||
| - | We transformed SCP+ToxR+FGFR (BBa_K569014) and ctx+GFP (BBa_J07011) into DH5a E.coli cells using two antibiotics: chloramphenicol and kanamycin. We selected a single colony and grew it in LB with chloramphenicol and kanamycin overnight at 37C and 220rpm. We made a 1:20 dilution of the sample and then grew it to a predetermined O.D. When the O.D. was reached induced with various amounts of heparin and fibroblast growth factor (FGF)and let it grow. We took measurements using fluorescence microscopy. | + | We transformed SCP+ToxR+FGFR (BBa_K569014) and ctx+GFP (BBa_J07011) into DH5a E.coli cells using two antibiotics: chloramphenicol and kanamycin. We selected a single colony and grew it in LB with chloramphenicol and kanamycin overnight at 37C and 220rpm. We made a 1:20 dilution of the sample and then grew it to a predetermined O.D. When the O.D. was reached induced with various amounts of heparin and fibroblast growth factor (FGF) and let it grow. We took measurements using fluorescence microscopy. |
<br><br> | <br><br> | ||
All images were taken with Olympus IX81 automated inverted microscope specially equipped for live cell imaging. The filter set we used is: 470/40x (excitation) and 525/50m (emission) for GFP. Data collection and processing was performed by the SlideBook software. | All images were taken with Olympus IX81 automated inverted microscope specially equipped for live cell imaging. The filter set we used is: 470/40x (excitation) and 525/50m (emission) for GFP. Data collection and processing was performed by the SlideBook software. | ||
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<img src="https://static.igem.org/mediawiki/2011/0/0e/SCP_1_hour.png" width="720"> | <img src="https://static.igem.org/mediawiki/2011/0/0e/SCP_1_hour.png" width="720"> | ||
<br><br> | <br><br> | ||
| - | We transformed PyeaR+ToxR+FGFR (BBa_K569013) and ctx+GFP (BBa_J07011) into DH5a E. | + | We transformed PyeaR+ToxR+FGFR (BBa_K569013) and ctx+GFP (BBa_J07011) into DH5a E.coli cells using two antibiotics: chloramphenicol and kanamycin. We selected a single colony and grew it in LB with chloramphenicol and kanamycin overnight at 37C and 220rpm. We made a 1:20 dilution of the sample and then grew it to a predetermined O.D. When the O.D. was reached we added potassium nitrate to a 10mM concentration and allowed the sample to grow. After 2 hours we induced with various amounts of heparin and fibroblast growth factor (FGF)and let it grow. We took measurements using fluorescence microscopy. |
<br><br> | <br><br> | ||
All images were taken with Olympus IX81 automated inverted microscope specially equipped for live cell imaging. The filter set we used is: 470/40x (excitation) and 525/50m (emission) for GFP. Data collection and processing was performed by the SlideBook software. | All images were taken with Olympus IX81 automated inverted microscope specially equipped for live cell imaging. The filter set we used is: 470/40x (excitation) and 525/50m (emission) for GFP. Data collection and processing was performed by the SlideBook software. | ||
<br><br> | <br><br> | ||
| - | Our results show | + | Our results show increased baseline expression of the CTX promoter and possible (but not substancial) increase in fluorescence when induced. |
<br><br> | <br><br> | ||
<img src="https://static.igem.org/mediawiki/2011/0/06/PyeaR_1_hour.png" width="720"> | <img src="https://static.igem.org/mediawiki/2011/0/06/PyeaR_1_hour.png" width="720"> | ||
Revision as of 23:39, 28 September 2011
Results
We transformed SCP+ToxR+FGFR (BBa_K569014) and ctx+GFP (BBa_J07011) into DH5a E.coli cells using two antibiotics: chloramphenicol and kanamycin. We selected a single colony and grew it in LB with chloramphenicol and kanamycin overnight at 37C and 220rpm. We made a 1:20 dilution of the sample and then grew it to a predetermined O.D. When the O.D. was reached induced with various amounts of heparin and fibroblast growth factor (FGF) and let it grow. We took measurements using fluorescence microscopy.All images were taken with Olympus IX81 automated inverted microscope specially equipped for live cell imaging. The filter set we used is: 470/40x (excitation) and 525/50m (emission) for GFP. Data collection and processing was performed by the SlideBook software.
We transformed PyeaR+ToxR+FGFR (BBa_K569013) and ctx+GFP (BBa_J07011) into DH5a E.coli cells using two antibiotics: chloramphenicol and kanamycin. We selected a single colony and grew it in LB with chloramphenicol and kanamycin overnight at 37C and 220rpm. We made a 1:20 dilution of the sample and then grew it to a predetermined O.D. When the O.D. was reached we added potassium nitrate to a 10mM concentration and allowed the sample to grow. After 2 hours we induced with various amounts of heparin and fibroblast growth factor (FGF)and let it grow. We took measurements using fluorescence microscopy.
All images were taken with Olympus IX81 automated inverted microscope specially equipped for live cell imaging. The filter set we used is: 470/40x (excitation) and 525/50m (emission) for GFP. Data collection and processing was performed by the SlideBook software.
Our results show increased baseline expression of the CTX promoter and possible (but not substancial) increase in fluorescence when induced.
