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SEPTEMBER, 8TH
PLATE
EXPERIMENT DESCRIPTION
Motivation
The experiment was performed to measure the 3OC6-HSL concentration due to the synthesis of homoserine-lactone by 3OC6-HSL generators in saturation phase of the cultures growth.
Methods
According to Protocol 3 the experiment was performed in LB on the following signalling parts:
- <partinfo>BBa_J61002</partinfo> - <partinfo>BBa_K300030</partinfo> (wiki name: I14)
- <partinfo>BBa_J61002</partinfo> - <partinfo>BBa_K300028</partinfo> (wiki name: I15)
- <partinfo>BBa_J61002</partinfo> - <partinfo>BBa_K300029</partinfo> (wiki name: I16)
- <partinfo>BBa_J61002</partinfo> - <partinfo>BBa_K300025</partinfo> (wiki name: I17)
- <partinfo>BBa_J61002</partinfo> - <partinfo>BBa_K300026</partinfo> (wiki name: I18)
- <partinfo>BBa_J61002</partinfo> - <partinfo>BBa_K300027</partinfo> (wiki name: I19)
- <partinfo>pSB4C5</partinfo> - <partinfo>BBa_K300030</partinfo> (wiki name: I14-4C5)
- <partinfo>pSB4C5</partinfo> - <partinfo>BBa_K300028</partinfo> (wiki name: I15-4C5)
- <partinfo>pSB4C5</partinfo> - <partinfo>BBa_K300029</partinfo> (wiki name: I16-4C5)
- <partinfo>pSB4C5</partinfo> - <partinfo>BBa_K300025</partinfo> (wiki name: I17-4C5)
- <partinfo>pSB4C5</partinfo> - <partinfo>BBa_K300026</partinfo> (wiki name: I18-4C5)
- <partinfo>pSB4C5</partinfo> - <partinfo>BBa_K300027</partinfo> (wiki name: I19-4C5)
- <partinfo>pSB1A2</partinfo> - <partinfo>BBa_B0031</partinfo> (negative control)
- <partinfo>pSB1AK3</partinfo> - <partinfo>BBa_K300009</partinfo> (negative control)
The light-blue wells refer to the same cultures grown over night; as the results were the same of the cultures grown for about six hours, from now on we have followed only the steps shown in Protocol 3.
RESULTS
It was possible to evaluate some of the 3OC6-HSL concentrations:
| Part | 3OC6-HSL concentration [uM]
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| <partinfo>BBa_J61002</partinfo> - <partinfo>BBa_K300030</partinfo> | ~0.7
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| <partinfo>BBa_J61002</partinfo> - <partinfo>BBa_K300028</partinfo> | ~0.04
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| <partinfo>BBa_J61002</partinfo> - <partinfo>BBa_K300025</partinfo> | ~0.09
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Because supernatants were not properly diluted it was not possible to measure some of the 3OC6-HSL concentrations; a more accurate evaluation of 3OC6-HSL was performed in the next test.
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