Team:Warsaw/Calendar-Main/3 October 2008

From 2008.igem.org

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<h3>Preparation of BioBricks</h3>
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<h3>Preparation of <a href=http://partsregistry.org/wiki/index.php?title=Part:pSB1A3>pSB1A3</a>+Omp_</h3>
<h4>Michał K.</h4>
<h4>Michał K.</h4>
<ol><li><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>+Omp_).</li>
<ol><li><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>+Omp_).</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>  
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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>
 +
<h4>Michał K.</h4>
 +
<ol>  
<li><a href="https://2008.igem.org/Wiki/Team:Warsaw/protocols#digest">Digest</a> of <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pET15b%2BOmpA-omega>pET15b+OmpA_omega</a> with XbaI (Tango buffer). DNA ends <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#blunting">blunting</a> with Klenow fragment (3 hr). </li>
<li><a href="https://2008.igem.org/Wiki/Team:Warsaw/protocols#digest">Digest</a> of <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pET15b%2BOmpA-omega>pET15b+OmpA_omega</a> with XbaI (Tango buffer). DNA ends <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#blunting">blunting</a> with Klenow fragment (3 hr). </li>
<li> Gel electrophoresis and <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#DNA_isolation_from_agarose_gel">gel-out</a> of proper band (<a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pET15b%2BOmpA-omega>pET15b+OmpA_omega</a> without XbaI - 6500 bp).</li>
<li> Gel electrophoresis and <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#DNA_isolation_from_agarose_gel">gel-out</a> of proper band (<a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pET15b%2BOmpA-omega>pET15b+OmpA_omega</a> without XbaI - 6500 bp).</li>

Revision as of 00:29, 27 October 2008

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Preparation of pSB2K3 + linker_alpha (BBa_K103009)

Piotr

  1. Transformation of TOP10 with ligation pSB2K3 + linker_alpha (BBa_K103009).
  2. Plating on LB with kanamycin.

Preparation of pSB2K3 + linker_omega (BBa_K103013)

Piotr

  1. Transformation of TOP10 with ligation pSB2K3 + linker_omega (BBa_K103013).
  2. Plating on LB with kanamycin.

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

Piotr

  1. Transformation of TOP10 with ligation pACYC177 + OmpA-linker-omega-linker (BBa_K103016).
  2. Plating on LB with kanamycin.

Preparation of pSB1A3+Omp_

Michał K.

  1. Isolation of plasmid from culture inoculated on previous day (pSB1A3+Omp_).
  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 pET15b+OmpA_omega with XbaI (Tango buffer). DNA ends blunting with Klenow fragment (3 hr).
  2. Gel electrophoresis and gel-out of proper band (pET15b+OmpA_omega without XbaI - 6500 bp).

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