Team:UNIPV-Pavia/Material Methods/Measurements/Tecan/test10settembre bis

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<html><p align="center"><font size="5"><b>19° TEST</b></font><br><br><font size="4">[SEPTEMBER, 10TH]</font></p></html><hr><br>
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==PLATE==
==PLATE==
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==EXPERIMENT DESCRIPTION==
==EXPERIMENT DESCRIPTION==
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==RESULTS (?)==
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===Motivation===
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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.
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===Methods===
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According to [[Team:UNIPV-Pavia/Parts/Characterization#Microplate reader experiments for self-inducible promoters - Protocol #3|Protocol 3]] the experiment was performed in LB on the following signalling parts:
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* <partinfo>BBa_J61002</partinfo> - <partinfo>BBa_K300030</partinfo> (wiki name: I14)
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* <partinfo>BBa_J61002</partinfo> - <partinfo>BBa_K300028</partinfo> (wiki name: I15)
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* <partinfo>BBa_J61002</partinfo> - <partinfo>BBa_K300029</partinfo> (wiki name: I16)
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* <partinfo>BBa_J61002</partinfo> - <partinfo>BBa_K300025</partinfo> (wiki name: I17)
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* <partinfo>BBa_J61002</partinfo> - <partinfo>BBa_K300026</partinfo> (wiki name: I18)
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* <partinfo>BBa_J61002</partinfo> - <partinfo>BBa_K300027</partinfo> (wiki name: I19)
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* <partinfo>pSB4C5</partinfo> - <partinfo>BBa_K300030</partinfo> (wiki name: I14-4C5)
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* <partinfo>pSB4C5</partinfo> - <partinfo>BBa_K300028</partinfo> (wiki name: I15-4C5)
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* <partinfo>pSB4C5</partinfo> - <partinfo>BBa_K300029</partinfo> (wiki name: I16-4C5)
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* <partinfo>pSB4C5</partinfo> - <partinfo>BBa_K300025</partinfo> (wiki name: I17-4C5)
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* <partinfo>pSB4C5</partinfo> - <partinfo>BBa_K300026</partinfo> (wiki name: I18-4C5)
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* <partinfo>pSB4C5</partinfo> - <partinfo>BBa_K300027</partinfo> (wiki name: I19-4C5)
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* <partinfo>pSB1A2</partinfo> - <partinfo>BBa_B0031</partinfo> (negative control)
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* <partinfo>pSB1AK3</partinfo> - <partinfo>BBa_K300009</partinfo> (negative control)
 +
 
 +
Supernatants were properly diluted in order to evaluate the production of 3OC6-HSL of the low copy number parts.
 +
 
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==RESULTS==
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The 3OC6-HSL concentrations of several cultures were evaluated:
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[[Image:UNIPV_Pavia_Serie110.png| 620px | center ]]
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[[Image:UNIPV_Pavia_Serie210.png| 620px | center ]]
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[[Image:UNIPV_Pavia_Serie310.png| 620px | center ]]
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Concentrations of 3OC6-HSL were obtained multiplying the values read on the graphics for the corresponding dilution factor.
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{|border='1' align='center'
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|| Part || 3OC6-HSL concentration [uM]
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|-
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|| <partinfo>BBa_J61002</partinfo> - <partinfo>BBa_K300030</partinfo> || ~0.4
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|-
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|| <partinfo>BBa_J61002</partinfo> - <partinfo>BBa_K300028</partinfo> || ~0.02
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|-
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|| <partinfo>BBa_J61002</partinfo> - <partinfo>BBa_K300025</partinfo> || ~0.1
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|-
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|| <partinfo>BBa_J61002</partinfo> - <partinfo>BBa_K300027</partinfo> || ~0.002
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|-
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|| <partinfo>pSB4C5</partinfo> - <partinfo>BBa_K300030</partinfo> || ~0.005
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|-
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|| <partinfo>pSB4C5</partinfo> - <partinfo>BBa_K300028</partinfo> || ~0.002
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|-
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|| <partinfo>pSB4C5</partinfo> - <partinfo>BBa_K300025</partinfo> || ~0.003
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|}
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[[Team:UNIPV-Pavia/Material_Methods/Measurements|MEASUREMENTS]]
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[[Team:UNIPV-Pavia/Material_Methods/Measurements/Tecan|Tecan Test]]
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Latest revision as of 10:27, 23 October 2010
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SEPTEMBER, 10TH


PLATE


UNIPV Pavia piastra10settembre.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)

Supernatants were properly diluted in order to evaluate the production of 3OC6-HSL of the low copy number parts.

RESULTS

The 3OC6-HSL concentrations of several cultures were evaluated:


UNIPV Pavia Serie110.png


UNIPV Pavia Serie210.png


UNIPV Pavia Serie310.png

Concentrations of 3OC6-HSL were obtained multiplying the values read on the graphics for the corresponding dilution factor.

Part 3OC6-HSL concentration [uM]
<partinfo>BBa_J61002</partinfo> - <partinfo>BBa_K300030</partinfo> ~0.4
<partinfo>BBa_J61002</partinfo> - <partinfo>BBa_K300028</partinfo> ~0.02
<partinfo>BBa_J61002</partinfo> - <partinfo>BBa_K300025</partinfo> ~0.1
<partinfo>BBa_J61002</partinfo> - <partinfo>BBa_K300027</partinfo> ~0.002
<partinfo>pSB4C5</partinfo> - <partinfo>BBa_K300030</partinfo> ~0.005
<partinfo>pSB4C5</partinfo> - <partinfo>BBa_K300028</partinfo> ~0.002
<partinfo>pSB4C5</partinfo> - <partinfo>BBa_K300025</partinfo> ~0.003




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