6 August 2013

From 2013.igem.org

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Running of an agarose gel

  • Running the gel for digests of samples 5.1, 10.1, 10.2 and 10.3 from the previous day.
  • expectation-7kb band, confirming the presence of the limonene biobrick (5kb) and chloramphenicol backbone (2kb)

Igem single dig 060813.jpg

  • The gel shows the presence of 7kb bands for both digests of samples 10.1, 10.2 and the 5.1 sample, which was expected.
  • No bands appeared for samples 5.1.
  • For samples 10.3 there is a 2kb band, indicating that with the ligation, the chloroamphenicol circulized.

Making chloroamphenicol agar plates

  • Making additional plates
    • 400ml of agar
      • Melt in microwave
      • 3mins on low -> shake
      • Repeat
      • 2mins on low -> shake
      • Repeat
    • Add 800ul of chloroamphenicol at a concentration of 25 ul/ml
    • Pour out onto 20 petri dishes
    • Leave to set on bench

Plating out samples

  • Plating out samples 10.1, 10.2, 10.3 and 5.1 onto chloroamphenicol plates.
  • Grow overnight at 37C