Team:Warsaw/Calendar-Main/31 July 2008
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<li>Lysis buffer was added separately to pellet and supernatant, then samples were boiled for 10 min. </li> | <li>Lysis buffer was added separately to pellet and supernatant, then samples were boiled for 10 min. </li> | ||
<li>Lysates loaded on 12% poliacrylamide gel (amount relating to 100 μl of OD=1.0 culture).</li> | <li>Lysates loaded on 12% poliacrylamide gel (amount relating to 100 μl of OD=1.0 culture).</li> | ||
- | <li>Gel stained with Coomassie Blue. Optimal induction conditions chosen (Fig. 1 and Fig. 2).</li> | + | <li>Gel stained with Coomassie Blue. Optimal induction conditions chosen (Fig. 1 and Fig. 2).</li></ol> |
Revision as of 08:16, 25 October 2008
Purification of proteins: Z-alpha and Z-omegaPiotr, Emilia
Cloning of omega_A DNA fragment to pACYC177+OmpA_alphaMichał K.
Cloning of truncated fragment of protein AMichał K.
The arrow shows place of our overexpressed protein: 1. 22°C 0.5 mM IPTG 2. 22°C 0.1 mM IPTG 3. 22°C 1 mM IPTG 4. Marker 5. negative control 6. 37°C 0.1 mM IPTG 7. 37°C 0.5 mM IPTG 8. 37°C 1 mM IPTG Fig. 2. Pellets and supernatants from Z-alpha induction with various IPTG concentrations The arrow shows place of our overexpressed protein: 1. Marker 2. pellet negative control 3. supernatant negative control 4. pellet 37°C 0.1 mM IPTG 5. supernatant 37°C 0.1 mM IPTG 6. sonicate 22°C 0.1 mM IPTG 7. sonicate 22°C 0.5 mM IPTG 8. pellet 37°C 0.5 mM IPTG 9. supernatant 37°C 0.5 mM IPTG
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