Team:UNIPV-Pavia/Material Methods/Measurements/Tecan/test8settembre

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Revision as of 09:49, 23 October 2010
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SEPTEMBER, 8TH


PLATE


UNIPV Pavia piastra8settembre.png


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

UNIPV Pavia Serie18.png

It was possible to evaluate some of the 3OC6-HSL concentrations:

Part 3OC6-HSL concentration [uM]
<partinfo>BBa_J61002</partinfo> - <partinfo>BBa_K300030</partinfo> ~0.7
<partinfo>BBa_J61002</partinfo> - <partinfo>BBa_K300028</partinfo> ~0.04
<partinfo>BBa_J61002</partinfo> - <partinfo>BBa_K300025</partinfo> ~0.09

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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Tecan Test

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