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Team:Calgary/Notebook/Protocols/catalase
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TITLE=Catalase Assay| | TITLE=Catalase Assay| | ||
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| - | + | <p> The purpose of this protocol is to test the ability of KatG-LAA to detoxify hydrogen peroxide. This was important, as catalase will be encorporated into the final sulfur circuit as an optimization feature. | |
<ol> | <ol> | ||
| - | <li>Grow cultures of <i>E. coli J04500-katG+LAA</i> and <i>J04500</i> overnight at 37*C in LB with appropriate antibiotics.</li> | + | <li><a href="https://2012.igem.org/Team:Calgary/Notebook/Protocols/onculture">Grow cultures</a> of <i>E. coli J04500-katG+LAA</i> and <i>J04500</i> overnight at 37*C in LB with appropriate antibiotics.</li> |
<li>The following evening, innoculate 20 uL into 3 mL LB with varying concentrations of hydrogen peroxide (0 mM, 1 mM, 5 mM, 10 mM) and grow overnight.</li> | <li>The following evening, innoculate 20 uL into 3 mL LB with varying concentrations of hydrogen peroxide (0 mM, 1 mM, 5 mM, 10 mM) and grow overnight.</li> | ||
| - | <li>The following morning, observe culture turbidity.</li> | + | <li>The following morning, observe and record culture turbidity.</li> |
</ol> | </ol> | ||
Latest revision as of 21:55, 26 October 2012
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Catalase Assay
The purpose of this protocol is to test the ability of KatG-LAA to detoxify hydrogen peroxide. This was important, as catalase will be encorporated into the final sulfur circuit as an optimization feature.
- Grow cultures of E. coli J04500-katG+LAA and J04500 overnight at 37*C in LB with appropriate antibiotics.
- The following evening, innoculate 20 uL into 3 mL LB with varying concentrations of hydrogen peroxide (0 mM, 1 mM, 5 mM, 10 mM) and grow overnight.
- The following morning, observe and record culture turbidity.
