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

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Revision as of 09:52, 23 October 2010

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. All supernatants were diluted 1:100.

RESULTS

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.

Tecan Test

@@ Line 40: / Line 40: @@
 * <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 [[Team:UNIPV-Pavia/Parts/Characterization#Microplate reader experiments for self-inducible promoters - Protocol #3|Protocol 3]].
+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 [[Team:UNIPV-Pavia/Parts/Characterization#Microplate reader experiments for self-inducible promoters - Protocol #3|Protocol 3]]. All supernatants were diluted 1:100.
 ==RESULTS==