Team:Warsaw/Calendar-Main/31 July 2008
From 2008.igem.org
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<h4> Michał K.</h4> | <h4> Michał K.</h4> | ||
<p><ol> | <p><ol> | ||
- | <li> <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#chemotransform">Transformation</a> of <i>E. coli</i> <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#top">TOP10</a> strain with ligation products (<a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pACYC177%2BompA-deltaA-alpha>pACYC177+OmpA_alpha + | + | <li> <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#chemotransform">Transformation</a> of <i>E. coli</i> <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#top">TOP10</a> strain with ligation products (<a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pACYC177%2BompA-deltaA-alpha>pACYC177+OmpA_alpha + ΔA</a> and <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pACYC177%2BOmpA-deltaA-omega>pACYC177+OmpA_omega + ΔA</a>). </li> |
<li> Transformants plating on LB + kanamycin. </li></ol></p> | <li> Transformants plating on LB + kanamycin. </li></ol></p> | ||
Revision as of 18:39, 26 October 2008
Cloning of truncated fragment of protein A (ΔA)Michał K.
Cloning of omega_ΔA DNA fragment to pACYC177+OmpA_alphaMichał K.
1-4. digested plasmids pACYC177 + OmpA-omega-deltaA-alpha 5. Marker Purification of proteins: Z-alpha and Z-omegaPiotr, Emilia
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. 3. 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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