Team:Warsaw/Calendar-Main/7 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 plasmid from culture 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>
<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/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 plasmid with EcoRI and PstI (Orange buffer). Gel electrophoresis - no proper clones found a
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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 - no proper clones found
<a href="https://2008.igem.org/Team:Warsaw/Calendar-Main/7_October_2008#fig2">Fig. 2</a>.</li>
<a href="https://2008.igem.org/Team:Warsaw/Calendar-Main/7_October_2008#fig2">Fig. 2</a>.</li>
<li>Inoculation of few more colonies which grown on plate with transformation: <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> to liquid LB + kanamycin. </li></ol>
<li>Inoculation of few more colonies which grown on plate with transformation: <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> to liquid LB + kanamycin. </li></ol>

Revision as of 21:03, 28 October 2008

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Preparation of linker_alpha (BBa_K103009)

Michał K., Piotr

  1. Isolation of plasmid from culture inoculated on previous day (pSB2K3 + linker_alpha (BBa_K103009)).
  2. Control digest of isolated plasmid with EcoRI and PstI (Orange buffer). Gel electrophoresis - no proper clones found.
  3. Inoculation of few more colonies which grown on plate with transformation: pSB2K3 + linker_alpha (BBa_K103009) to liquid LB + kanamycin.

Preparation of linker_omega (BBa_K103013)

Michał K., Piotr

  1. Isolation of plasmid from culture inoculated on previous day (pSB2K3 + linker_omega (BBa_K103013)).
  2. Control digest of isolated plasmid with EcoRI and PstI (Orange buffer). Gel electrophoresis - no proper clones found Fig. 1.
  3. Inoculation of few more colonies which grown on plates with transformation: pSB2K3 + linker_omega (BBa_K103013) to liquid LB + kanamycin.
Fig. 1.Control EcoRI/PstI digest of pSB2K3+linker_omega (BBa_K103013)
1. Marker
2-17. Control digests of pSB2K3+linker_omega (BBa_K103013)

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

Michał K., Piotr

  1. Isolation of plasmid from culture inoculated on previous day (pACYC177 + OmpA-linker-omega-linker (BBa_K103016)).
  2. Control digest of isolated plasmid with EcoRI and PstI (Orange buffer). Gel electrophoresis - no proper clones found Fig. 2.
  3. Inoculation of few more colonies which grown on plate with transformation: pACYC177 + OmpA-linker-omega-linker (BBa_K103016) to liquid LB + kanamycin.
Fig. 2. Control digests of pACYC177+OmpA-linker-omega-linker (BBa_K103016)
1. Marker
2.-11. Control digests of pACYC177+OmpA-linker-omega-linker (BBa_K103016)
12. Marker

Preparation of vector for pT7 constructs

Piotr

  1. Transformation of TOP10 with selfligation of pET15b+OmpA_omega (with removed XbaI site).
  2. Plating on LB with ampicillin.

Preparation of OmpA_linker_omega_linker under Plac (BBa_K103018)

Piotr

  1. Transformation of TOP10 with ligations pSB2K3 + BBa_K103018 (without internal EcoRI site).
  2. Plating on LB with kanamycin.

Preparation of AID(BBa_K103001)

Michał K., Piotr

  1. Ligation of DNA fragments: pSB1A3+ AID(BBa_K103001) (from 1 October).
  2. Transformation of TOP10 with above ligation.
  3. Plating on LB with ampicillin.

Preparation of AID under pBAD/araC (BBa_K103002)

Piotr

  1. Digest of pMPMT5+AID with EcoRI (EcoRI buffer). DNA ends blunting with Klenow fragment.
  2. Overnight ligation of pMPMT5+AID to remove EcoRI site.

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