Team:Warsaw/Calendar-Main/31 July 2008
From 2008.igem.org
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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.</li> | + | <li>Gel stained with Coomassie Blue. Optimal induction conditions chosen (Fig. 1 and Fig. 2).</li> |
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<p> | <p> | ||
<ol><li> <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#plasmid_DNA_isolation">Isolation of plasmids</a> from cultures inocluated on <a href="https://2008.igem.org/Team:Warsaw/Calendar-Main/31_July_2008">previous day</a>. </li> | <ol><li> <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#plasmid_DNA_isolation">Isolation of plasmids</a> from cultures inocluated on <a href="https://2008.igem.org/Team:Warsaw/Calendar-Main/31_July_2008">previous day</a>. </li> | ||
- | <li> Control <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#digest">digest</a> of isolated plasmids with BamHI and SacI (BamHI buffer). </li><li> Gel electrophoresis - proper | + | <li> Control <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#digest">digest</a> of isolated plasmids with BamHI and SacI (BamHI buffer). </li><li> Gel electrophoresis - proper clones founded. </li></ol> |
</p> | </p> | ||
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<li> Transformants plating on LB + kanamycin. </li></ol></p> | <li> Transformants plating on LB + kanamycin. </li></ol></p> | ||
+ | <img src="https://static.igem.org/mediawiki/2008/e/e7/July_31_st.jpg" width=300 /><var><b>Z-omega induction with various IPTG concentrations<br> | ||
+ | The arrow shows place of our overexpressed protein</b>: | ||
+ | 1. 22°C 0,5 mM IPTG<br> | ||
+ | 2. 22°C 0,1 mM IPTG<br> | ||
+ | 3. 22°C 1 mM IPTG<br> | ||
+ | 4. Marker<br> | ||
+ | 5. negative control<br> | ||
+ | 6. 37°C 0,1 mM IPTG<br> | ||
+ | 7. 37°C 0,5 mM IPTG<br> | ||
+ | 8. 37°C 1 mM IPTG<br></var> | ||
</html> | </html> | ||
Revision as of 15:18, 24 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
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