IGEM:The University of Sheffield/2009/Notebook/Sheffield iGEM 2010/2010/08/24

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Tuesday 24th August:

  • Transformations failed.
  • Ordered competent DH5-alpha e.coli from Bioline.
  • Set up an overnight restriction digest of pgaA promoter with EcoR1 and Pst1 restriction enzymes:
  Plasmid- 10μL
  Pst1 - 2μL
  EcoR1 - 2μL
  EcoR1 buffer - 2μL
  BSA (x20) - 1μL
  H2O - 3μL


  • Ran an electrophoresis gel[[1]] of plasmids pACYC duet-1 and pCDF. Results shown bellow:

UVP00418.JPG

  • The first three wells from the bottom were filled with the cut pACYC duet-1.
  • The three wells between the two ladders contain the cut pCDF.
  • pACYC duet-1 was extracted from the gel.[[2]]