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

From 2010.igem.org

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




Tecan Test