Difference between revisions of "Part:BBa K137076:Experience"
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===Applications of BBa_K137076=== | ===Applications of BBa_K137076=== | ||
| − | *Strain: This part will only work in a catalase deficient strain of E. coli. Our testing strain, JI377, is DE(katE, katG, ahp). | + | *Strain: This part will only work in a catalase deficient strain of E. coli. Our testing strain, JI377, is DE(katE, katG, ahp). No H<sub>2</sub>O<sub>2</sub> is produced in a catalase proficient strain, such as DH10B. |
*Culture conditions: Saturated LB culture backdiluted 1:100 into SOC +1mM IPTG. Let shake at 37C until the culture reaches an OD600 = 0.8. Bring culture to 10nM AHL and let shake at 37 C for 3 hrs. Pellet cells and assay supernatant for hydrogen peroxide. | *Culture conditions: Saturated LB culture backdiluted 1:100 into SOC +1mM IPTG. Let shake at 37C until the culture reaches an OD600 = 0.8. Bring culture to 10nM AHL and let shake at 37 C for 3 hrs. Pellet cells and assay supernatant for hydrogen peroxide. | ||
*Results: Under these conditions the supernatant had ~50 uM H<sub>2</sub>O<sub>2</sub> prior to induction and ~200 uM ~200uM H<sub>2</sub>O<sub>2</sub> 3hrs post induction with AHL. | *Results: Under these conditions the supernatant had ~50 uM H<sub>2</sub>O<sub>2</sub> prior to induction and ~200 uM ~200uM H<sub>2</sub>O<sub>2</sub> 3hrs post induction with AHL. | ||
Latest revision as of 07:18, 14 August 2008
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Applications of BBa_K137076
- Strain: This part will only work in a catalase deficient strain of E. coli. Our testing strain, JI377, is DE(katE, katG, ahp). No H2O2 is produced in a catalase proficient strain, such as DH10B.
- Culture conditions: Saturated LB culture backdiluted 1:100 into SOC +1mM IPTG. Let shake at 37C until the culture reaches an OD600 = 0.8. Bring culture to 10nM AHL and let shake at 37 C for 3 hrs. Pellet cells and assay supernatant for hydrogen peroxide.
- Results: Under these conditions the supernatant had ~50 uM H2O2 prior to induction and ~200 uM ~200uM H2O2 3hrs post induction with AHL.
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