Team:Heidelberg/Notebook/Sensing Group/Notebook/3rdweek

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Monday, 08/18/2008

  • PCR again for LuxQ-1 and LuxQ-2 for Fusion constructs. Different Mg concentrations from 1% to 4% were used. For LuxQ-2 another PCR was run using as template V. harveyi colonies
PCR of LuxQ-1 and LuxQ-2 for subsequent fusion PCR
  • Fusion-PCR for Fusion-1 and Fusion-2, each 4 tubes (2 µl Tar mixed with 1 µl LuxQ)
  • preparation of O/N culture of LuxS and pDK48 transformed cells for subsequent miniprep
  • digestion of pDK48 with NcoI/NdeI (NEBuffer 4)
  • digestion of pTrc99alpha and LuxQ with BamHI/NcoI (NEBuffer 3 + BSA) and gel extraction

Tuesday, 08/19/2008

  • Miniprep of pDK48 and LuxS
  • gel of Fusion constructs and gelextraction of pDK48 from yesterday
  • afterwards another gel was run to purify (get rid of the primers used) the Fusion constructs via gel extraction
Fusion constructs and pDK48 gel extraction

Wednesday, 08/20/2008 - Friday, 08/22/2008

  • nothing important to report

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