Team:Warsaw/Calendar-Main/25 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-Z-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-Z-omega>pACYC177+OmpA_Z_omega</a></h3><h4>Michał K.</h4>
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<p> Eight colonies from each plate (<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.</p>
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<p> Eight colonies from each plate (<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>) were inoculated to liquid LB with kanamycin.</p>
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Tests for ampicillin resistance given by protein added to medium

Piotr

  1. Purification of pACYC177+OmpA_alpha from antibiotic resistance contamination:
    Inoculation of E. coli TOP10 careing pACTC177+OmpA_alpha to liquin LB with kanamycin and with kanamycin and ampicylin.
  2. Inoculation of TOP10 carrying pACYC177+OmpA_A_alpha and pACYC177+OmpA-alpha to LB with inductor (0.25 mM IPTG) and kanamycin.

Cloning of protein A DNA to pET15b+OmpA-alpha plasmid

Antoni

  1. Transformation of E. coli TOP10 strain with friday's ligation products.
  2. Transformants plating on LB + ampicillin.

Cloning of alpha to pACYC177+OmpA_A_omega and pACYC177+OmpA_Z_omega

Michał K.

Eight colonies from each plate (pACYC177+OmpA_alpha_A_omega and pACYC177+OmpA_alpha_Z_omega) were inoculated to liquid LB with kanamycin.