Team:NTU-Singapore/Notebook/20 June 2008

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Contents

Friday 20 June

Morning:

  • Miniprep for pLacI, followed by DNA concentrating using QiAQuick PCR purification kit.

Afternoon:

  • Transformation and cell cloning 4 empty plasmids from Biobrick registry:
    • [http://partsregistry.org/wiki/index.php?title=Part:pSB1A3 pSB1A3]
    • [http://partsregistry.org/wiki/index.php?title=Part:pSB1AC3 pSB1AC3]
    • [http://partsregistry.org/wiki/index.php?title=Part:pSB1AK3 pSB1AK3]
    • [http://partsregistry.org/wiki/index.php?title=Part:pSB1AT3 pSB1AT3]
  • Transformation and cloning of LacI-GFP into LuxS knockout bacteria.
  • Make LsrA competent cells. Then transform 0.3 ul LacI-GFP plasmid into these cells.

Evening:

  • Gel extraction (using QiAQuick Gel extraction Kit)==>Ligation (using T4 DNA ligase) ==> Transformation for the following 4 samples:
    • E7 Insert - Empty plasmid vector (from GFP)
    • SupD Insert - Empty plasmid vector (from GFP)
    • T7ptag Insert - Empty plasmid vector (from GFP)
    • GFP insert - pFE vector (for characterization of Fe promoter)
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