Team:Groningen/Labwork/14 August 2013

From 2013.igem.org

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<h2>Inne</h2>
<h2>Inne</h2>
Made an inoculation of <i>E.coli</i> from Utah, first in a Erlenmeyer with 200 ml LB medium, 500 uL Cm (5mg/mL) and 2 mL of previously made culture.
Made an inoculation of <i>E.coli</i> from Utah, first in a Erlenmeyer with 200 ml LB medium, 500 uL Cm (5mg/mL) and 2 mL of previously made culture.
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<br>Then pipetted 3mL of this into a test-tube to grow overnight at 37C.  
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<br>Then pipetted 3mL of this into a test-tube to grow overnight at 37C.
 +
<br>The motility assay didn't provide the results we were looking for. The Negative control of tubes with pipet tips was contaminated, the other tubes did show colonies in small dots in the agar of the test tubes. This is probably from bacteria floating up when the agar was poured. 
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Revision as of 12:47, 15 August 2013

Mirjam

Ligation reaction of ΔCheY and ΔDes (with help of Chaline)
Transformation to B. subtilis ΔDes transformed in ΔCheY and ΔCheY into ΔDes.
Inoculation of Pdes-CheY-GFP in LB medium with ampicillin (although strange colonies grew on the negative control plate). The colonies didn't grew in liquid LB medium.

Sander

made 3 inoculations of the cheY knockout and 3 inoculations of wild type bacillus in 0,4% agar with LB medium to test motility.

Chaline

Did a restriction on CheY down and Des up with EcoRI and heat inactivated the samples.
Run the samples over gel and the bands are visible.
Measured the concentration of CheY down and Des up.

Inne

Made an inoculation of E.coli from Utah, first in a Erlenmeyer with 200 ml LB medium, 500 uL Cm (5mg/mL) and 2 mL of previously made culture.
Then pipetted 3mL of this into a test-tube to grow overnight at 37C.
The motility assay didn't provide the results we were looking for. The Negative control of tubes with pipet tips was contaminated, the other tubes did show colonies in small dots in the agar of the test tubes. This is probably from bacteria floating up when the agar was poured.