Team:Warsaw/Calendar-Main/20 August 2008

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<h3>Cloning of alpha to <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pACYC177%2BompA-A-omega>pACYC177+OmpA_A_omega</a> and <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pACYC177%2BompA-A-omega>pACYC177+OmpA_Z_omega</a></h3><h4>Michał K.</h4>
<h3>Cloning of alpha to <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pACYC177%2BompA-A-omega>pACYC177+OmpA_A_omega</a> and <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pACYC177%2BompA-A-omega>pACYC177+OmpA_Z_omega</a></h3><h4>Michał K.</h4>
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<p> Colonies from both plates(<a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pACYC177%2BompA-alpha-A-omega>pACYC177+OmpA_alpha_A_omega</a> and <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pACYC177%2BOmpA-alpha-Z-omega>pACYC177+OmpA_alpha_Z_omega</a>) was inoculated to liquid LB with kanamycin (only two colonies grown on each plate).</p>
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<p> Colonies from both plates(<a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pACYC177%2BompA-alpha-A-omega>pACYC177+OmpA_alpha_A_omega</a> and <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pACYC177%2BOmpA-alpha-Z-omega>pACYC177+OmpA_alpha_Z_omega</a>) was inoculated to liquid LB with kanamycin (only two colonies have grown on each plate).</p>
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Tests for ampicillin resistance given by protein added to medium

Piotr

  1. Inoculation of all pACYC177-OmpA variants of constructs to LB + 0.25 mM IPTG, kanamycin, ampicillin (50 μg/ml) and 100 μg/ml of protein (Z-alpha and Z-omega; negative control without protein).
  2. Measurement of growth after 5 hours and on next day.

Cloning of protein A DNA to GeneArt plasmid

Antoni

Transformation of E. coli TOP10 strain with ligation products from previous day.

Cloning of alpha to pACYC177+OmpA_A_omega and pACYC177+OmpA_Z_omega

Michał K.

Colonies from both plates(pACYC177+OmpA_alpha_A_omega and pACYC177+OmpA_alpha_Z_omega) was inoculated to liquid LB with kanamycin (only two colonies have grown on each plate).