Team:Paris Bettencourt/Notebook/Trojan Horse/Wednesday 24th July.html

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Trojan Horse

24th July



Glycerol stock:
  • sT008: MGZ1, F+
  • sT009: XL1-Blue Litmus28i

Infectiveness characterization experiments
  • Dilute 1/100 MGZ1, F+ O/N culture in 5mL LB => wait OD(600) = 0.7
  • Centrifuge M13K07 4000rpm, 10 minutes
  • Retrieve surnageant and filter it with 0.2 filter
  • Once OD 0.7 is reached, make 4 tubes
    1. 1mL MGZ1,F+ + 100ul LB (not done, not enough cells left ofter OD test)
    2. 1mL MGZ1,F+ + 100ul surnageant diluted 1/100
    3. 1mL MGZ1,F+ + 100ul surnageant diluted 1/1000
    4. 1mL MGZ1,F+ + 100ul surnageant diluted 1/1000
  • Incube 37°C for 45 minutes
  • Serial dilute the tubes 10^-1, 10^-2, 10^-3, 10^-4, 10^-5, 10^-6
    • 4 times 6 tubes with 900uL LB, add 100ul of one previously made tube, mix and transfert 100ul to next tube.
  • Plate on LB (10^-6), Kan(10^-2 and 10^-3), Amp(10^-6), Kan and Amp(10^-2 and 10^-3)

Experiment fail due to manipulation error (wrong dilution of the supernageant and plating of wrong dilutions)

Start O/N culture of MGZ1,F+ (T008) + Tet 1/500
Start O/N culture of M13K07 (T007) + Amp and Kan 1/1000

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