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Team:Warsaw/Calendar-Main/29 July 2008
From 2008.igem.org
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<p>Inoculation OmpA_omega_A_alpha from various IPTG concentrations (in mmol/mL) (0, 0.1, 0.25, 0.5, 0.75, 1) into same IPTG concentrations, but with various ampicillin concentrations (in mmol/mL)(25, 50, 75, 100) in ratio: 1:50.</p> | <p>Inoculation OmpA_omega_A_alpha from various IPTG concentrations (in mmol/mL) (0, 0.1, 0.25, 0.5, 0.75, 1) into same IPTG concentrations, but with various ampicillin concentrations (in mmol/mL)(25, 50, 75, 100) in ratio: 1:50.</p> | ||
| - | <center><h4>Measurement of bacterial culture growth (OD) in the evening:</h4> </center> | + | <center><h4 style="text-align: center">Measurement of bacterial culture growth (OD) in the evening:</h4> </center> |
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</table> | </table> | ||
<p> | <p> | ||
| - | Inoculation of OmpA_omega_A_alpha into various IPTG concentrations: 0, 0.1, 0.25, 0.5, 0.75 mmol/mL ( | + | Inoculation of OmpA_omega_A_alpha into various IPTG concentrations: 0, 0.1, 0.25, 0.5, 0.75 mmol/mL (replication is necessary)</p> |
<h3>Checking OmpA_omega_A_alpha and OmpA_A_alpha expression<br> | <h3>Checking OmpA_omega_A_alpha and OmpA_A_alpha expression<br> | ||
Revision as of 14:34, 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 | 1.558 | 1.469 | 1.587 | 1.49 | 1.566 | 1.311 |
| 50 | 1.425 | 1.435 | 1.524 | 1.055 | 0.920 | 0.935 |
| 75 | 1.09 | 0.989 | 1.447 | 0.971 | 0.951 | 0.992 |
| 100 | 0.09 | 0.685 | 1.378 | 1.078 | 0.977 | 0.992 |
Inoculation of OmpA_omega_A_alpha into various IPTG concentrations: 0, 0.1, 0.25, 0.5, 0.75 mmol/mL (replication is necessary)
Checking OmpA_omega_A_alpha and OmpA_A_alpha expression
Piotr
- Spinning
- Suspending
- Adding of lysis buffer
- Boiling
- Putting into poliacrylamide gel
- Transfer onto nitrocellulose
- Blocking
- Anti-A antibody binding
- Washing
- Anti-rabbit antibody binding
- Developing with BCIP and NBT
Michał L., Ewa, Marcin
- Separate transformant colonies (transformation from previous day) inoculated to liquid LB with kanamycin.
- Inoculation of pACYC177+OmpA_alpha and pACYC177+OmpA_omega - liquid LB with kanamycin.
