Team:Warsaw/Calendar-Main/14 October 2008

From 2008.igem.org

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<h3>Preparation of BioBricks</h3>
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<h3>Preparation of pLac_OmpA_alpha</h3>
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<h4>Michał K.</h4>
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<h3></h3>
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<h4>Piotr</h4>
<h4>Piotr</h4>
<ol>
<ol>
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<li> <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#chemotransform">Transformation</a> of <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#top10">TOP10</a> with overnight ligations.</li>
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<li> <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#chemotransform">Transformation</a> of <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#top10">TOP10</a> with overnight ligation - <a href=http://partsregistry.org/Part:pSB2K3>pSB2K3</a>_pLac_OmpA_ + alpha2.</li>
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<li> Tranformants planting on LB with kanamycin - 3kb(?????)+ link_alpha, 3kb(????)_pLac_OmpA_ + alpha2 and RFP(?????) + AraC+pBAD+AID on LB with ampicillin. </li>
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<li> Tranformants planting on LB with kanamycin. </li>
</ol>
</ol>

Revision as of 12:06, 27 October 2008

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Preparation of linker_alpha (BBa_K103009)

Piotr

  1. Transformation of TOP10 with overnight ligation - pSB2K3 + linker_alpha (BBa_K103009).
  2. Tranformants plating on LB with kanamycin.

Preparation of OmpA-linker-omega-linker (BBa_K103016)

Michał K.

  1. Isolation of plasmid from culture inoculated on previous day - pACYC177 + OmpA-linker-omega-linker (BBa_K103016).
  2. Control digest of isolated plasmids with EcoRI and PstI (Orange buffer). Gel electrophoresis - still no proper clones found.

Preparation of alpha_linker under PT7 (BBa_K103019)

Michał K.

Isolation of plasmid from culture inoculated on previous day - pSB1A3 carrying alpha_linker under PT7 (BBa_K103019).

Preparation of pT7_omega_link

Michał K.

  1. Isolation of plasmid from culture inoculated on previous day (pSB2K3 + pT7_omega_link).
  2. Control digest of isolated plasmids with EcoRI and PstI (Orange buffer). Gel electrophoresis - proper clones found.

Preparation of AraC+pBAD+AID

Michał K.

  1. Transformation of TOP10 with overnight ligation - pSB1A3 + AraC+pBAD+AID.
  2. Tranformants planting on LB with ampicillin.

Preparation of pLac_OmpA_alpha

Piotr

  1. Transformation of TOP10 with overnight ligation - pSB2K3_pLac_OmpA_ + alpha2.
  2. Tranformants planting on LB with kanamycin.

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