8 August 2013

From 2013.igem.org

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Making samples for DNA sequencing

Making chloroamphenicol/IPTG plates

  • 400ml of agar
    • Boil for 3min x2
    • Boil for 2min x2
  • Cool to 40C
  • Add 800ul of chloroamphenicol at 25ug/ml
  • Add 400ul IPTG at 0.15mM
  • Makes 20 petri dishes

Streaked out samples of the limonene/pSB1C3 ligated plasmid

  • Streaked samples 5.1, 10.1, 10.2 and 10.3 onto the chloroamphenicol/IPTG plates
  • IPTG was used to induce the lac operon, and therefore induce limonene synthesis
  • Plates were labelled using special symbols by only one team member to ensure double-blinded test

Made polystyrene strips

--> include video here

Overnight culture preparation

  • To make glycerol broths, so sample can be frozen and preserved
  • Use four universal tubes, add:
    • 4ml Luria Broth
    • 8ul chloroamphenicol
    • Add bacteria from the original plates (sample 5.1, 10.1, 10.2 and 10.3 into a single tube each respectively)
*Each sample was also re-streaked on a chloroamphenicol plate to maintain current stock.