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Team:Osaka/Tests
From 2012.igem.org
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== Tests == | == Tests == | ||
=== Damage tolerance assay === | === Damage tolerance assay === | ||
| - | To measure the DNA damage tolerance conferred by each part, we used antibacterial agents as a source of DNA damage and then assayed the survival rates. Transformed ''E. coli'' was exposed to antibacterial agents and then incubated for 2 hours. Cells were plated on agar plates at different dilutions | + | To measure the DNA damage tolerance conferred by each part, we used antibacterial agents as a source of DNA damage and then assayed the survival rates. Transformed ''E. coli'' was exposed to antibacterial agents and then incubated for 2 hours. Cells were plated on agar plates at different dilutions and air dried. 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 tolerance parts tested were as follows: | The tolerance parts tested were as follows: | ||
Revision as of 09:37, 11 September 2012
Tests
Damage tolerance assay
To measure the DNA damage tolerance conferred by each part, we used antibacterial agents as a source of DNA damage and then assayed the survival rates. Transformed E. coli was exposed to antibacterial agents and then incubated for 2 hours. Cells were plated on agar plates at different dilutions and air dried. 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 tolerance parts tested were as follows:
Parts containing one gene each
- CDS: PprI, PprA, PprM or RecA
Parts containing two genes
- CDS1+2: PprI+RecA, PprA+RecA, PprM+RecA, PprI+PprA, PprI+PprM, PprA+PprM
Discussion
Single-gene parts
Two-gene combinations
Conclusion
SOS promoter assay
We assayed the promoter of the SOS gene RecA ([http://partsregistry.org/wiki/index.php?title=Part:BBa_J22106 J22106]). To measure the DNA damage detection, we used antibacterial agents as a source of DNA damage. To quantitatively and accurately evaluate the promoter activity, dual luciferase assay method was employed. Transformed E. coli was exposed to antibacterial agents and then incubated for 2 hours. For details check the Protocols page.
