Team:Warsaw/Calendar-Main/20 August 2008

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<h3>Tests for ampicillin resistance given by protein added to medium</h3><h4>Piotr</h4>
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<ol><li>Inoculation of all OmpA variants of constructs to LB + 0.25 mM IPTG, kanamycin, ampicillin (50 μg/ml) and 100 μg of protein (A-alpha, Z-alpha and Z-omega; negative control without protein).</li>
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<li>Measurement of growth after 5 hours and on next day.</li></ol>
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<h3>Cloning of protein A DNA to GeneArt plasmid</h3><h4>Antoni</h4>
<h3>Cloning of protein A DNA to GeneArt plasmid</h3><h4>Antoni</h4>

Revision as of 21:49, 19 October 2008

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Tests for ampicillin resistance given by protein added to medium

Piotr

  1. Inoculation of all OmpA variants of constructs to LB + 0.25 mM IPTG, kanamycin, ampicillin (50 μg/ml) and 100 μg of protein (A-alpha, 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 grown on each plate).