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
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
Fusion constructs and pDK48 gel extraction

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

  • nothing important to report

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