Team:Evry/Notebook/CellCharacterization/Antibiotic test/08-18-2014

From 2014.igem.org

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Tests of antibiotics' stocks - Results(2)

Plates LB Agar only LB+Kan LB+Erm (1:100)
E.coli X 0 0

New stocks are ready for antibiotics' tests on Pseudovribrio

Launch of antibiotics's tests in liquid M9 1X
For each tested antibiotic we made the following range of concentrations:

0 1:500 1:1000 1:50 000 1:100 000

We made these range two time to have a replicate and we added a controle tube without antibiotic for each bacteria. Each tube is made like in the following table in Falcon tubes (15mL):


/ 0 1:500 1:1000 1:50 000 1:100 000
Medium+agar 3mL 3mL 3mL 3mL 3mL
Antibiotic 0 6µL of stock 3µL of stock 6µL of stock diluted 1/100 3µL of stock diluted 1/100
Pre-culture 300µL 300µL 300µL 300µL 300µL

For each range, we test Pseudovibrio denitrificans but also E.Coli and E.Coli tranformed with a speicific resistance cassette (NM:The E.Coli strain for the control case is the same strain that is tranformed for the cassette efficiency test). The initial OD at 600nm of each bacteria has been mesured to after be able to calculate the ΔOD at 600nm, during three days.


/ Pseudo Bl21 DH5a Top 10 DH5a pyr+KanR DH5a+CamR Top 10+ErmR
OD init 0.02 0.795 0.873 0.693 0.408 0.687 0.319

Plates for antibiotics's tests
For each tested antibiotic we made the same range of concentrations than in liquid. We also made these range two time to have a replicate and we added a controle plate without antibiotic on which we showed all our tested bacteria. Plates will be checked during three days. We made these same ranges of tested antibiotic's concentrations for three media:

  • MB agar 1X
  • MB agar 0.5X
  • M9 agar 1X

Each plate is made like in the following table:


/ 0 1:500 1:1000 1:50 000 1:100 000
Medium+agar 20mL 20mL 20mL 20mL 20mL
Antibiotic 0 40µL of stock 20µL of stock 40µL of stock diluted 1/100 20µL of stock diluted 1/100

Aug 18