Team:Warsaw/Calendar-Main/23 September 2008

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(Difference between revisions)
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<h3>Preparation of BioBricks</h3>
<h3>Preparation of BioBricks</h3>
<h4>Michał K.</h4>
<h4>Michał K.</h4>
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<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.</li>
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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.</li>
<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). No proper colonies found.</li>
<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). No proper colonies found.</li>

Revision as of 21:22, 21 October 2008

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Mutagenesis of protein A

Paweł

  1. PNK phosporylation of mutagenesis products for 30 min at 37°C.
  2. PNK heat-inactivated by incubation at 75°C for 15 min.
  3. T4 ligase added and incubated for 1 h at room temperature.
  4. T4 ligase inactivated by incubation at 55°C for 20 min, then 5U of DpnI added and incubated at 37°C for 3 hours.
  5. After DpnI treatment mutagenesis products transformed into TOP10 and plated on LB + kanamycin plates.

Preparation of BioBricks

Michał K.

  • Isolation of plasmids from cultures inoculated on previous day.
  • Control digest of isolated plasmids with EcoRI and PstI (Orange buffer). No proper colonies found.


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