IGEM:The University of Sheffield/2009/Notebook/Sheffield iGEM 2010/2010/09/14

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Tuesday 14th September

  • Carried out plasmid mini preps of all the successful transformations using the overnight cultures from yesterday.[[1]]
  • Set up an overnight digestions of the promoters using the following method:
  • Promoter digest:
 Plasmid - 10μL
 Pst1 - 2μL
 EcoR1 - 2μL
 EcoR1 buffer - 2μL
 BSA (x20) - 1μL
 H2O - 3μL
  • Set up an overnight digestion of part BBa_J04450, using the following method:
  • BBa_J04450 digest:
 Plasmid - 10μL
 Pst1 - 2μL
 EcoR1 - 2μL
 EcoR1 buffer - 2μL
 BSA (x20) - 1μL
 H2O - 3μL
  • Part BBa_J04450 is a RFP contained in a pSB1C3 plasmid and is also Cm resistant. This digestion is therefore to remove the RFP so that this plasmid can be used for submission of our promoter-GFP biobricks to iGEM.




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