Team:Warsaw/Calendar-Main/14 October 2008

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<h3>Preparation of pT7_alpha_link</h3>
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<h4>Michał K.</h4>
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<p><a href=https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#plasmid_DNA_isolation>Isolation</a> of plasmid from culture inoculated on previous day - <A href=http://partsregistry.org/wiki/index.php?title=Part:pSB1A3>pSB1A3</a> carrying pT7_alpha fragment.</p>

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Preparation of pSB2K3 + 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 pACYC177 + 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 pT7_alpha_link

Michał K.

Isolation of plasmid from culture inoculated on previous day - pSB1A3 carrying pT7_alpha fragment.

Preparation of BioBricks

Michał K.

  1. Isolation of plasmid from culture inoculated on previous day (5kb (??????) + pT7_omega_link, RFP(?????) + pT7_alpha and 3kb (??????) + OmpA_omega_link).
  2. Control digest of isolated plasmids (5kb (??????) + pT7_omega_link and 3kb (??????) + OmpA_omega_link) with EcoRI and PstI (Orange buffer). Gel electrophoresis - still no proper clones found for 3kb (??????) + OmpA_omega_link ligation but we found good clones for (5kb (??????) + pT7_omega_link.

Piotr

  1. Transformation of TOP10 with overnight ligations.
  2. Tranformants planting on LB with kanamycin - 3kb(?????)+ link_alpha, 3kb(????)_pLac_OmpA_ + alpha2 and RFP(?????) + AraC+pBAD+AID on LB with ampicillin.