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Team:Warsaw/Calendar-Main/30 July 2008
From 2008.igem.org
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<p>Transformation of <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#rosetta">Rosettas</a> with omega_A_alpha and A_alpha.</p> | <p>Transformation of <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#rosetta">Rosettas</a> with omega_A_alpha and A_alpha.</p> | ||
| + | <h4>Michał K.:</h4> | ||
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| + | <li>Isolation of plasmids from cultures inocluated on previous day.</li> | ||
| + | <li>Control <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#digest">digest</a> of isolated plasmids (pACYC177+omega_deltaA) with HindIII and SacI (7 from 8 colonies confirmed). </li></ol></p> | ||
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Revision as of 14:25, 9 October 2008
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Checking if OmpA_omega_A_alpha gives ampicillin resistance
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| ampicillin concentration (μg/mL): | IPTG concentration (mmol/mL): | |||||
|---|---|---|---|---|---|---|
| 0 | 0.1 | 0.25 | 0.5 | 0.75 | 1 | |
| 25 | 1.054 | 1.154 | 1.051 | 0.99 | 1.096 | 0.896 |
| 50 | 0.94 | 0.891 | 1.123 | 0.847 | 0.924 | 0,81 |
| 75 | 0.5 | 0.63 | 0.743 | 0.782 | 0.631 | 0.64 |
| 100 | 0.02 | 0.396 | 0.563 | 0.678 | 0.602 | 0.611 |
Conclusion: we obtained the best induction using 0.5 - 0.25 mM IPTG. Best ampicillin concentration is 50 - 75 μg/ml.
Checking if degradation of fusion with OmpA is a result of the activity of lon and iompt proteases. (present in top10)
Piotr
Test was conducted in E.coli Rosetta strain expressing omp_omega_A_alfa (with and without induction) and omp_A_alfa.
Transformation of Rosettas with omega_A_alpha and A_alpha.
Michał K.:
- Isolation of plasmids from cultures inocluated on previous day.
- Control digest of isolated plasmids (pACYC177+omega_deltaA) with HindIII and SacI (7 from 8 colonies confirmed).
