Team:Warsaw/Calendar-Main/7 October 2008

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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.</li>
<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.</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>
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<a name="fig1"><img src="https://static.igem.org/mediawiki/2008/0/0e/Traw_05_10_2008.jpg"></a>
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<var><b>Fig. 22.</b>Traw_05_10_2008.jpg </var>

Revision as of 23:58, 27 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.
  3. Inoculation of few more colonies which grown on plates with transformation: pSB2K3 + linker_omega (BBa_K103013) to liquid LB + kanamycin.

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.
  3. Inoculation of few more colonies which grown on plate with transformation: pACYC177 + OmpA-linker-omega-linker (BBa_K103016) to liquid LB + kanamycin.
Fig. 22.Traw_05_10_2008.jpg

Preparation of vector for pT7 constructs

Piotr

  1. Transformation of TOP10 with selfligation of pET15b+OmpA_omega without XbaI.
  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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