Team:Warsaw/Calendar-Main/2 October 2008

From 2008.igem.org

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<li> Confirmed transformant colonies inoculated to liquid LB with ampicillin.</li>
<li> Confirmed transformant colonies inoculated to liquid LB with ampicillin.</li>
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<h3>Preparation of <a href=http://partsregistry.org/wiki/index.php?title=Part:pSB1A3>pSB1A3</a>+<a href=http://partsregistry.org/wiki/index.php?title=Part:BBa_K103004>Z(BBa_K103004)</a></h3>
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<h4>Michał K.</h4>
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<ol><li><a href=https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#plasmid_DNA_isolation>Isolation</a> of plasmids from culture inoculated on previous day (<a href=http://partsregistry.org/wiki/index.php?title=Part:pSB1A3>pSB1A3</a>+<a href=http://partsregistry.org/wiki/index.php?title=Part:BBa_K103004>Z(BBa_K103004)</a>).</li>
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<li>Control <a href=https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#digest>digest</a> of isolated plasmid with EcoRI and PstI (Orange buffer). Gel electrophoresis - proper clones found.</li></ol>
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<h3>Preparation of BioBricks</h3>
<h3>Preparation of BioBricks</h3>
<h4>Michał K.</h4>
<h4>Michał K.</h4>
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<ol><li><a href=https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#plasmid_DNA_isolation>Isolation</a> of plasmids from culture inoculated on previous day (<a href=http://partsregistry.org/wiki/index.php?title=Part:pSB1A3>pSB1A3</a>+<a href=http://partsregistry.org/wiki/index.php?title=Part:BBa_K103004>Z(BBa_K103004)</a>).</li>
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<ol>
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<li>Control <a href=https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#digest>digest</a> of isolated plasmid with EcoRI and PstI (Orange buffer). Gel electrophoresis - proper clones found.</li>
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<li> <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#digest">Digest</a> of pLac_OmpA_omega fragment with EcoRI and BcuI (BamHI buffer). </li>
<li> <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#digest">Digest</a> of pLac_OmpA_omega fragment with EcoRI and BcuI (BamHI buffer). </li>
<li>Gel elctrophoresis and <a href="https://2008.igem.org/Wiki/Team:Warsaw/protocols#DNA_isolation_from_agarose_gel">gel-out</a> of proper band - 1200 bp. </li>
<li>Gel elctrophoresis and <a href="https://2008.igem.org/Wiki/Team:Warsaw/protocols#DNA_isolation_from_agarose_gel">gel-out</a> of proper band - 1200 bp. </li>

Revision as of 10:24, 27 October 2008

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Preparation of pSB2K3 + linker_alpha (BBa_K103009)

Michał K.

Overnight ligation of isolated DNA fragments: pSB2K3 + linker_alpha (BBa_K103009).

Preparation of pSB2K3 + linker_omega (BBa_K103013)

Michał K.

Overnight ligation of isolated DNA fragments: pSB2K3 + linker_omega (BBa_K103013).

Preparation of pACYC177 + OmpA-linker-omega-linker (BBa_K103016)

Michał K.

Overnight ligation of DNA fragments isolated on 30 September : pACYC177 + OmpA-linker-omega-linker (BBa_K103016).

Preparation of pSB1A3+OmpA-linker (BBa_K103006)

Michał K.

  1. Colony PCR with OmpaL_N and OmpaP_link primers on colonies from plates with transformations pSB1A3+OmpA-linker (BBa_K103006) (annealing temperature - 55°C,45 s of elongation step).
  2. Gel electrophoresis.
  3. Confirmed transformant colonies inoculated to liquid LB with ampicillin.

Preparation of pSB1A3+Z(BBa_K103004)

Michał K.

  1. Isolation of plasmids from culture inoculated on previous day (pSB1A3+Z(BBa_K103004)).
  2. Control digest of isolated plasmid with EcoRI and PstI (Orange buffer). Gel electrophoresis - proper clones found.

Preparation of BioBricks

Michał K.

  1. Digest of pLac_OmpA_omega fragment with EcoRI and BcuI (BamHI buffer).
  2. Gel elctrophoresis and gel-out of proper band - 1200 bp.

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