Team:Warsaw/Calendar-Main/2 October 2008

From 2008.igem.org

(Difference between revisions)
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  primers on colonies from plates with transformations <a href=http://partsregistry.org/wiki/index.php?title=Part:pSB1A3>pSB1A3</a>+<a href=http://partsregistry.org/Part:BBa_K103006>OmpA-linker (BBa_K103006)</a> (annealing temperature - 55&deg;C,45 s of elongation step). </li>
  primers on colonies from plates with transformations <a href=http://partsregistry.org/wiki/index.php?title=Part:pSB1A3>pSB1A3</a>+<a href=http://partsregistry.org/Part:BBa_K103006>OmpA-linker (BBa_K103006)</a> (annealing temperature - 55&deg;C,45 s of elongation step). </li>
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<li> Gel electrophoresis(<a href="https://2008.igem.org/Team:Warsaw/Calendar-Main/2_October_2008#fig1">Fig. 1.</a>).</li>
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<li> Gel electrophoresis (<a href="https://2008.igem.org/Team:Warsaw/Calendar-Main/2_October_2008#fig1">Fig. 1.</a>).</li>
<li> Confirmed transformant colonies inoculated to liquid LB with ampicillin.</li>
<li> Confirmed transformant colonies inoculated to liquid LB with ampicillin.</li>
</ol>
</ol>
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<var><b>Fig. 1.</b>Colony PCR with OmpaL_N and OmpaP_link. <br>
<var><b>Fig. 1.</b>Colony PCR with OmpaL_N and OmpaP_link. <br>
1. Marker<br>
1. Marker<br>
-
4, 14, 19, 20, 24. Transformation confirmed. <br> </var>
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4, 14, 19, 20, 24. Proper bands visible. <br> </var>
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<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 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>
<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> Gel electrophoresis(<a href="https://2008.igem.org/Team:Warsaw/Calendar-Main/2_October_2008#fig2">Fig. 2.</a>).</li></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> Gel electrophoresis (<a href="https://2008.igem.org/Team:Warsaw/Calendar-Main/2_October_2008#fig2">Fig. 2.</a>).</li></ol>
<a name="fig2"><img src="https://static.igem.org/mediawiki/2008/6/6f/Traw_02_10_2008.jpg"></a>
<a name="fig2"><img src="https://static.igem.org/mediawiki/2008/6/6f/Traw_02_10_2008.jpg"></a>
<var><b>Fig. 2.</b>Control digest of pSB1A3+Z(BBa_K103004)<br>
<var><b>Fig. 2.</b>Control digest of pSB1A3+Z(BBa_K103004)<br>
1. Marker<br>
1. Marker<br>
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3. Clone carrying BBa_K103004</var>
+
3-5. Clones carrying BBa_K103004</var>

Revision as of 20:31, 28 October 2008

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Preparation of linker_alpha (BBa_K103009)

Michał K.

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

Preparation of linker_omega (BBa_K103013)

Michał K.

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

Preparation of 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 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 (Fig. 1.).
  3. Confirmed transformant colonies inoculated to liquid LB with ampicillin.
Fig. 1.Colony PCR with OmpaL_N and OmpaP_link.
1. Marker
4, 14, 19, 20, 24. Proper bands visible.

Preparation of 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 (
  3. Gel electrophoresis (Fig. 2.).
Fig. 2.Control digest of pSB1A3+Z(BBa_K103004)
1. Marker
3-5. Clones carrying BBa_K103004

Preparation of OmpA_linker_omega_linker under Plac (BBa_K103018)

Michał K.

  1. Digest of BBa_K103018 fragment with EcoRI and BcuI (BamHI buffer).
  2. Gel electrophoresis
  3. Gel electrophoresis (Fig. 3.) and gel-out of proper band - 1200 bp.
Fig. 3.Digest of OmpA_linker_omega_linker under Plac (BBa_K103018).
1. Marker
2. OmpA_linker_omega_linker under Plac (BBa_K103018)

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