Team:UNIPV-Pavia/Material Methods/Measurements/Tecan/test2settembre

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SEPTEMBER, 2ND


PLATE


UNIPV Pavia piastra2settembre.png


EXPERIMENT DESCRIPTION

Motivation

This experiment was performed in M9 to check bacterial growth and RFP rate synthesis in order to evaluate the R.P.U. (see Protocol) of the promoters from Anderson's collection in high copy and low copy number plasmids.

Methods

Accordign to Protocol 2 the experiment was performed in M9 on the following parts:

  • <partinfo>BBa_J23100</partinfo>
  • <partinfo>BBa_J23101</partinfo>
  • <partinfo>BBa_J23105</partinfo>
  • <partinfo>BBa_J23106</partinfo>
  • <partinfo>BBa_J23110</partinfo>
  • <partinfo>BBa_J23114</partinfo>
  • <partinfo>BBa_J23116</partinfo>
  • <partinfo>BBa_J23118</partinfo>
  • <partinfo>pSB4C5</partinfo> - <partinfo>BBa_J23100</partinfo> (wiki name: BBa_J23100_4C5)
  • <partinfo>pSB4C5</partinfo> - <partinfo>BBa_J23101</partinfo> (wiki name: BBa_J23101_4C5)
  • <partinfo>pSB4C5</partinfo> - <partinfo>BBa_J23105</partinfo> (wiki name: BBa_J23105_4C5)
  • <partinfo>pSB4C5</partinfo> - <partinfo>BBa_J23106</partinfo> (wiki name: BBa_J23106_4C5)
  • <partinfo>pSB4C5</partinfo> - <partinfo>BBa_J23110</partinfo> (wiki name: BBa_J23110_4C5)
  • <partinfo>pSB4C5</partinfo> - <partinfo>BBa_J23114</partinfo> (wiki name: BBa_J23114_4C5)
  • <partinfo>pSB4C5</partinfo> - <partinfo>BBa_J23116</partinfo> (wiki name: BBa_J23116_4C5)
  • <partinfo>pSB4C5</partinfo> - <partinfo>BBa_J23118</partinfo> (wiki name: BBa_J23118_4C5)
  • <partinfo>BBa_B0031</partinfo> (negative control)

RESULTS

UNIPV Pavia HCpromotersM9.png

In high copy number plasmids it is possible to identify 3 groups: strong promoters (<partinfo>Ba_J23100</partinfo> and <partinfo>BBa_J23101</partinfo>), medium strength promoters (in order <partinfo>BBa_J23110</partinfo> and <partinfo>BBa_J23118</partinfo>) and a group of weak promotrs.

Promoter R.P.U. (average) std
<partinfo>BBa_J23100</partinfo> 0.946 0.182
<partinfo>BBa_J23101</partinfo> 1 -
<partinfo>BBa_J23105</partinfo> 0.196 0.038
<partinfo>BBa_J23106</partinfo> 0.326 0.015
<partinfo>BBa_J23110</partinfo> 0.678 0.096
<partinfo>BBa_J23114</partinfo> 0.091 0.022
<partinfo>BBa_J23116</partinfo> 0.101 0.004
<partinfo>BBa_J23118</partinfo> 0.655 0.035
UNIPV Pavia LCpromotersM9.png

In low copy number plasmids there are the same 3 groups but <partinfo>BBa_J23118</partinfo> resulted stronger than <partinfo>BBa_J23110</partinfo>.

Promoter R.P.U. (average) std
<partinfo>pSB4C5</partinfo> - <partinfo>BBa_J23100</partinfo> 0.838 0.101
<partinfo>pSB4C5</partinfo> - <partinfo>BBa_J23101</partinfo> 1 -
<partinfo>pSB4C5</partinfo> - <partinfo>BBa_J23105</partinfo> 0.149 0.045
<partinfo>pSB4C5</partinfo> - <partinfo>BBa_J23106</partinfo> 0.286 0.033
<partinfo>pSB4C5</partinfo> - <partinfo>BBa_J23110</partinfo> 0.466 0.101
<partinfo>pSB4C5</partinfo> - <partinfo>BBa_J23114</partinfo> 0.053 0.048
<partinfo>pSB4C5</partinfo> - <partinfo>BBa_J23116</partinfo> 0.110 0.007
<partinfo>pSB4C5</partinfo> - <partinfo>BBa_J23118</partinfo> 0.714 0.154



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