Team:Warsaw/Calendar-Main/5 August 2008

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<ol><li><a href="https://2008.igem.org/Wiki/Team:Warsaw/protocols#digest">Digest</a> of pACYC177+OmpA_omega_deltaA_alpha (from 25 July) with BamHI and NotI. DNA ends blunting with Klenow fragment (3 hr). </li>
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<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 - 4300 bp. </li>
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<li><a href="https://2008.igem.org/Wiki/Team:Warsaw/protocols#ligation">Ligation</a> of isolated DNA fragment (1 hr). </li>
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<li><a href="https://2008.igem.org/Wiki/Team:Warsaw/protocols#chemotransform">Transformation</a>of E. coli <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#top10">TOP10</a> strain with ligation. </li>
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<li>Transformants plating on LB + kanamycin. </li></ol>
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<h3>Michał</h3>
<h3>Michał</h3>
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Revision as of 21:01, 11 October 2008

Gallery Bricks Notebook Team Project Home


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  1. Digest of pACYC177+OmpA_omega_deltaA_alpha (from 25 July) with BamHI and NotI. DNA ends blunting with Klenow fragment (3 hr).
  2. Gel elctrophoresis and gel-out of proper band - 4300 bp.
  3. Ligation of isolated DNA fragment (1 hr).
  4. Transformationof E. coli TOP10 strain with ligation.
  5. Transformants plating on LB + kanamycin.

Michał

  1. Isolation of plasmids from cultures inocluated on previous day.
  2. Control digest of isolated plasmids with BamHI and SacI.

Checking for presence of A on the cell membrane
Piotr

  1. Pouring bacteria by drops onto nitrocellulose
  2. Drying
  3. Blocking
  4. Anti-A antibody binding
  5. Washing
  6. Anti-rabbit antibody binding
  7. Developing with BCIP and NBT

[photo of the gel is to be placed here]


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