Team:Heidelberg/Notebook/Sensing Group/Notebook/8thweek

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Contents

Monday, 09/22/2008

  • test digestion of F1_LuxP_NdeI (F1 cloned in pDK48 with NdeI), F2_LuxP_NdeI, F1_LuxP_KpnI (F1 cloned in pDK48 with KpnI) and F2_LuxP_KpnI (F2 cloned in pDK48 with KpnI) with NcoI
Digestions of Fusion constructs with NcoI. Positive clones show two bands at 4308 and 2915 bp
  • Transformation of Fusion constructs (F1(NdeI)_LuxP_pDK48, F1(KpnI)_LuxP_pDK48 and F2(NdeI)_LuxP_pDK48) into MG1655 and HCB33 cells
  • Preparation of cultures for Maxipreps of Fusion constructs

Tuesday, 09/23/2008

  • Maxiprep of LuxP-Fusion constructs (F1(NdeI)_LuxP_pDK48, F1(KpnI)_LuxP_pDK48 and F2(NdeI)_LuxP_pDK48)
  • Cloning of Fusion_LuxP in pBAD33 with BamHI/PstI
  • digestions of Fusion constructs and pBAD33 as described in gel image descriptions
Digestion of Fusion constructs and pBAD33 with BamHI/PstI for subsequent cloning.
Digestion of Fusion constructs with NcoI and PstI to check for correct insert and Maxiprep digetions.


  • O/N culture of Fusion-LuxP-pDK48 in HCB33 and MG1655 in TB-Kan
  • Transformation of Fusion-LuxP-pBAD33 constructs into DH5a

Wednesday, 09/24/2008

  • Glycerol-Stock of Fusion1-LuxP-pDK48 and Fusion2-LuxP-pDK48
  • O/N culture of Fusion-LuxP-pDK48 in HCB33 and MG1655 in LB
  • O/N culture of LuxS and GFP Plasmid from Marcus
  • Re-Sequencing of Fusion-1 and Fusion-2 
Correct sequence for both Fusion constructs confirmed

Thursday, 09/25/2008

  • Miniprep of Fusion-LuxP-pDK48 constructs, LuxS, GFP Plasmid from Marcus
  • Competent Cells (Top-10, UU1250)
  • Picked colonies of Fusion-LuxP-pBAD33

Friday, 09/26/2008

Go to 9th week

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