Team:Warsaw/Calendar-Main/30 September 2008

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<li><a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#digest">Digest</a> of isolated plasmids with EcoRI and BcuI (BamHI buffer). </li>
<li><a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#digest">Digest</a> of isolated plasmids with EcoRI and BcuI (BamHI buffer). </li>
<li> <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#digest">Digest</a> of pACYC177+OmpA_omega_deltaA_alpha with BamHI and PstI (BamHI buffer). </li>
<li> <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#digest">Digest</a> of pACYC177+OmpA_omega_deltaA_alpha with BamHI and PstI (BamHI buffer). </li>
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<li><a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#removing">Dephosphorylation</a> (CIAP) of plasmids.</li>
<li>Gel elctrophoresis and <a href="https://2008.igem.org/Wiki/Team:Warsaw/protocols#DNA_isolation_from_agarose_gel">gel-out</a> of proper band: ??????. </li>  
<li>Gel elctrophoresis and <a href="https://2008.igem.org/Wiki/Team:Warsaw/protocols#DNA_isolation_from_agarose_gel">gel-out</a> of proper band: ??????. </li>  
<li><a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#digest">Digest</a> of omega_link fragment (PCR from previous day) with EcoRI and BcuI (BamHI buffer)</li>
<li><a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#digest">Digest</a> of omega_link fragment (PCR from previous day) with EcoRI and BcuI (BamHI buffer)</li>

Revision as of 12:47, 25 October 2008

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Mutagenesis of protein A

Paweł

Results of sequencing: unfortunately all sequences where wild-type.

Preparation of BioBricks

Michał K.

  1. Isolation of HQs(??????) plasmids.
  2. Digest of isolated plasmids with EcoRI and BcuI (BamHI buffer).
  3. Digest of pACYC177+OmpA_omega_deltaA_alpha with BamHI and PstI (BamHI buffer).
  4. Dephosphorylation (CIAP) of plasmids.
  5. Gel elctrophoresis and gel-out of proper band: ??????.
  6. Digest of omega_link fragment (PCR from previous day) with EcoRI and BcuI (BamHI buffer)
  7. Digest of OmpA_omega with BglII and PstI (Orange buffer).
  8. Clean-up of above digest reactions.
  9. Overnight digest of purified pLac_OmpA_omega fragment (from previous day) with EcoRI (EcoRI buffer). DNA ends blunting with Klenow fragment.

Piotr

  1. E. coli TOP10 transformation with overnight ligations (RFP(??????) + Z and RFP(????) + OmpA_link).
  2. Plating on LB + ampicillin.
  3. Inoculation of bacterias received from iGEM HQs ( ).