IGEM:The University of Sheffield/2009/Notebook/Sheffield iGEM 2010/2010/09/08

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Wednesday 8th September

  • Gel Electrophoresis [[1]] was repeated with the two differentlly cut plasmids containing Part BBa_E0040:

The Gel was set up as follows starting from the bottom: 1) Part BBa_E0040 cut with EcoR1 and Pst1. 2) Part BBa_E0040 cut with EcoR1 and Xpa1. 3) Ladder.

UVP00460.JPG

  • Both bands from each lane were removed using the gel extraction kit.[[2]]
  • A ligation was set up between the cholera system and the cut plasmid pACYC. Six different variations of the ligase constituents were used to see which produced the best results:
Substance(μL) Tube 1 Tube 2 Tube 3 Tube 4
pACYC 4 2 2 2
System 2 2 4 2
Ligase 1 1 1 0
Buffer 1 1 1 1
H2O 2 4 2 5