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Team:Osaka/Tests
From 2012.igem.org
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To measure the DNA damage tolerance conferred by each part, we used UV irradiation as a source of DNA damage and then assayed the survival rates. Transformed ''E. coli'' cells were plated on agar plates at different dilutions, air dried, and then exposed to different doses of UV radiation. Plates were wrapped with aluminum foil and incubated in the dark. Colony-forming units were scored after 16h incubation at 37°C. For detailed protocols, refer to the [https://2012.igem.org/Team:Osaka/Protocols Protocols page]. | To measure the DNA damage tolerance conferred by each part, we used UV irradiation as a source of DNA damage and then assayed the survival rates. Transformed ''E. coli'' cells were plated on agar plates at different dilutions, air dried, and then exposed to different doses of UV radiation. Plates were wrapped with aluminum foil and incubated in the dark. Colony-forming units were scored after 16h incubation at 37°C. For detailed protocols, refer to the [https://2012.igem.org/Team:Osaka/Protocols Protocols page]. | ||
| - | + | = The results will be up soon! = | |
=== SOS promoter assay === | === SOS promoter assay === | ||
<p>We assayed the promoter of the SOS gene RecA ([http://partsregistry.org/wiki/index.php?title=Part:BBa_J22106 J22106]), by attaching a lycopene biosynthesis gene cluster ([http://partsregistry.org/Part:BBa_K274100 K274100]) downstream as a reporter to yield the DNA damage detection device ([http://partsregistry.org/Part:BBa_K602013 K602013]). | <p>We assayed the promoter of the SOS gene RecA ([http://partsregistry.org/wiki/index.php?title=Part:BBa_J22106 J22106]), by attaching a lycopene biosynthesis gene cluster ([http://partsregistry.org/Part:BBa_K274100 K274100]) downstream as a reporter to yield the DNA damage detection device ([http://partsregistry.org/Part:BBa_K602013 K602013]). | ||
Transformed ''E. coli'' was exposed to UV light and then incubated for 2 hours. Lycopene as a reporter was extracted from cells with acetone. For details check the [https://2012.igem.org/Team:Osaka/Protocols Protocols page].</p> | Transformed ''E. coli'' was exposed to UV light and then incubated for 2 hours. Lycopene as a reporter was extracted from cells with acetone. For details check the [https://2012.igem.org/Team:Osaka/Protocols Protocols page].</p> | ||
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| + | = The results will be up soon! = | ||
Revision as of 05:57, 17 August 2012
Tests
Damage tolerance assay
To measure the DNA damage tolerance conferred by each part, we used UV irradiation as a source of DNA damage and then assayed the survival rates. Transformed E. coli cells were plated on agar plates at different dilutions, air dried, and then exposed to different doses of UV radiation. Plates were wrapped with aluminum foil and incubated in the dark. Colony-forming units were scored after 16h incubation at 37°C. For detailed protocols, refer to the Protocols page.
The results will be up soon!
SOS promoter assay
We assayed the promoter of the SOS gene RecA ([http://partsregistry.org/wiki/index.php?title=Part:BBa_J22106 J22106]), by attaching a lycopene biosynthesis gene cluster ([http://partsregistry.org/Part:BBa_K274100 K274100]) downstream as a reporter to yield the DNA damage detection device ([http://partsregistry.org/Part:BBa_K602013 K602013]). Transformed E. coli was exposed to UV light and then incubated for 2 hours. Lycopene as a reporter was extracted from cells with acetone. For details check the Protocols page.
= The results will be up soon! =
