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

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SEPTEMBER, 10TH

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)

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:

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

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

Tecan Test