Team:Warsaw/Calendar-Main/17 October 2008

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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 cultures inoculated on previous day <a href=http://partsregistry.org/Part:BBa_I739204>pACYC177</a> + <a href=http://partsregistry.org/Part:BBa_K103016>OmpA-linker-omega-linker (BBa_K103016)</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 cultures inoculated on previous day <a href=http://partsregistry.org/Part:BBa_I739204>pACYC177</a> + <a href=http://partsregistry.org/Part:BBa_K103016>OmpA-linker-omega-linker (BBa_K103016)</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 plasmids with EcoRI and PstI (Orange buffer). Gel electrophoresis - no proper clones found.</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 plasmids with EcoRI and PstI (Orange buffer). Gel electrophoresis - no proper clones found <a href="https://2008.igem.org/Team:Warsaw/Calendar-Main/17_October_2008#fig1">Fig. 1</a>.</li></ol>
<a name="fig1"><img src="https://static.igem.org/mediawiki/2008/7/75/Traw_17_10_2008.jpg"></a>
<a name="fig1"><img src="https://static.igem.org/mediawiki/2008/7/75/Traw_17_10_2008.jpg"></a>
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<var><b>Fig. 26.</b>traw_17_10_2008.jpg </var>
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<var><b>Fig. 1.</b>EcoRI/PstI digests of plasmids pACYC177+OmpA-linker-omega-linker (BBa_K103016).<br>
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1. Marker <br>
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2-9. EcoRI/PstI digests of pACYC177+OmpA-linker-omega-linker (BBa_K103016)<br></var>
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<ol>
<ol>
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<li>Gel electrophoresis of over night digest (<A href=http://partsregistry.org/wiki/index.php?title=Part:pSB1A3>pSB1A3</a> carrying  <a href=http://partsregistry.org/Part:BBa_K103019>alpha_linker under PT7 (BBa_K103019)</a>) and <a href="https://2008.igem.org/Wiki/Team:Warsaw/protocols#DNA_isolation_from_agarose_gel">gel-out</a> of proper band - 800 bp <a href="https://2008.igem.org/Team:Warsaw/Calendar-Main/17_October_2008#fig1">Fig. 1</a>. </li>
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<li>Gel electrophoresis of over night digest (<A href=http://partsregistry.org/wiki/index.php?title=Part:pSB1A3>pSB1A3</a> carrying  <a href=http://partsregistry.org/Part:BBa_K103019>alpha_linker under PT7 (BBa_K103019)</a>) and <a href="https://2008.igem.org/Wiki/Team:Warsaw/protocols#DNA_isolation_from_agarose_gel">gel-out</a> of proper band - 800 bp. <a href="https://2008.igem.org/Team:Warsaw/Calendar-Main/17_October_2008#fig2">Fig. 2</a>. </li>
<li><a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#ligation">Ligation</a> of isolated DNA fragments: <a href=http://partsregistry.org/wiki/index.php?title=Part:pSB2K3>pSB2K3</a> (from <a href=https://2008.igem.org/Team:Warsaw/Calendar-Main/30_September_2008>30 September</a>) + <a href=http://partsregistry.org/Part:BBa_K103019>alpha_linker under PT7 (BBa_K103019)</a>.</li>
<li><a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#ligation">Ligation</a> of isolated DNA fragments: <a href=http://partsregistry.org/wiki/index.php?title=Part:pSB2K3>pSB2K3</a> (from <a href=https://2008.igem.org/Team:Warsaw/Calendar-Main/30_September_2008>30 September</a>) + <a href=http://partsregistry.org/Part:BBa_K103019>alpha_linker under PT7 (BBa_K103019)</a>.</li>
<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 above ligation.</li>
<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 above ligation.</li>
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<a name="fig2"><img src="https://static.igem.org/mediawiki/2008/e/e5/Go_17_10_2008.jpg"></a>
<a name="fig2"><img src="https://static.igem.org/mediawiki/2008/e/e5/Go_17_10_2008.jpg"></a>
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<var><b>Fig. 2.</b>EcoRI/BcuRI digests of BBa_K103019</var>
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<var><b>Fig. 2.</b>EcoRI/BcuRI digests of BBa_K103019.<br>
1. Marker<br>
1. Marker<br>
2. EcoRI/BcuI digest of BBa_K103019<br></var>
2. EcoRI/BcuI digest of BBa_K103019<br></var>

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

Michał K.

  1. Isolation of plasmids from cultures 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 - no proper clones found Fig. 1.
Fig. 1.EcoRI/PstI digests of plasmids pACYC177+OmpA-linker-omega-linker (BBa_K103016).
1. Marker
2-9. EcoRI/PstI digests of pACYC177+OmpA-linker-omega-linker (BBa_K103016)

Preparation of alpha_linker under PT7 (BBa_K103019)

Michał K., Piotr

  1. Gel electrophoresis of over night digest (pSB1A3 carrying alpha_linker under PT7 (BBa_K103019)) and gel-out of proper band - 800 bp. Fig. 2.
  2. Ligation of isolated DNA fragments: pSB2K3 (from 30 September) + alpha_linker under PT7 (BBa_K103019).
  3. Transformation of TOP10 with above ligation.
  4. Plating on LB with kanamycin.
Fig. 2.EcoRI/BcuRI digests of BBa_K103019.
1. Marker
2. EcoRI/BcuI digest of BBa_K103019

Preparation of AID under pBAD/araC (BBa_K103002)

Piotr

  1. Transformation of TOP10 with overnight ligation - pSB1A3 + AID under pBAD/araC (BBa_K103002).
  2. Plating on LB with ampicillin.

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