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Team:Evry/Notebook/CellCharacterization/Antibiotic test/08-18-2014
From 2014.igem.org
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| - | <u>Tests of antibiotics' stocks - Results(2)</u> <br> | + | <u><big><b><FONT COLOR=#003333>Tests of antibiotics' stocks - Results(2)</font></b></big></u> <br> |
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New stocks are ready for antibiotics' tests on Pseudovribrio<br> | New stocks are ready for antibiotics' tests on Pseudovribrio<br> | ||
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| - | <u>Launch of antibiotics's tests in liquid M9 1X</u> | + | <u>Launch of antibiotics's tests in liquid M9 1X</u> <br> |
For each tested antibiotic we made the following range of concentrations:<br> | For each tested antibiotic we made the following range of concentrations:<br> | ||
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We made these range two time to have a replicate and we added a controle tube without antibiotic for each bacteria. | 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):<br> | + | Each tube is made like in the following table in Falcon tubes (15mL):<br></p> |
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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). | 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.<br> | + | The initial OD at 600nm of each bacteria has been mesured to after be able to calculate the ΔOD at 600nm, during three days.<br></p> |
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<u>Plates for antibiotics's tests</u> <br> | <u>Plates for antibiotics's tests</u> <br> | ||
For each tested antibiotic we made the same range of concentrations than in liquid. | 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 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:<br> | + | We made these same ranges of tested antibiotic's concentrations for three media:<br></p> |
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<li>MB agar 1X | <li>MB agar 1X | ||
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<li>M9 agar 1X | <li>M9 agar 1X | ||
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| - | Each plate is made like in the following table:<br> | + | <p> |
| + | Each plate is made like in the following table:<br></p> | ||
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<span class="cd-date">Aug 18</span> | <span class="cd-date">Aug 18</span> | ||
Latest revision as of 22:26, 17 October 2014
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
