Team:Warsaw/Calendar-Main/25 August 2008

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<h3>Cloning of protein A DNA to <A href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pET15b%2BOmpA-alpha>pET15b+Omp-alpha</a> plasmid</h3><h4>Antoni</h4>
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<h3>Cloning of protein A DNA to <A href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pET15b%2BOmpA-alpha>pET15b+OmpA_alpha</a> plasmid</h3><h4>Antoni</h4>
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Revision as of 18:07, 24 October 2008

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Tests for ampicillin resistance given by protein added to medium

Piotr

  1. Purification OmpA_alpha of 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 OmpA_A_alpha and 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) was inoculated to liquid LB with kanamycin.

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