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

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<html><p align="center"><font size="5"><b>17° TEST</b></font><br><br><font size="4">[SEPTEMBER, 2ND]</font></p></html><hr><br>
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<html><p align="center"><font size="5"><b>SEPTEMBER, 2ND</b></font></p></html><hr><br>
==PLATE==
==PLATE==
<br>
<br>
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===Motivation===
===Motivation===
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This experiment was performed in M9 to check bacterial growth and RFP rate synthesis in order to evaluate the R.P.U. () of the promoters from Anderson's collection in high copy and low copy number plasmids.
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This experiment was performed in M9 to check bacterial growth and RFP rate synthesis in order to evaluate the R.P.U. ([[Team:UNIPV-Pavia/Parts/Characterization#Data analysis for RPU evaluation|see Protocol]]) of the promoters from Anderson's collection in high copy and low copy number plasmids.
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==RESULTS (?)==
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===Methods===
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Accordign to [[Team:UNIPV-Pavia/Parts/Characterization#Microplate reader experiments for constitutive promoters (R.P.U. evaluation) - Protocol #2|Protocol 2]] the experiment was performed in M9 on the following parts:
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* <partinfo>BBa_J23100</partinfo>
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* <partinfo>BBa_J23101</partinfo>
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* <partinfo>BBa_J23105</partinfo>
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* <partinfo>BBa_J23106</partinfo>
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* <partinfo>BBa_J23110</partinfo>
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* <partinfo>BBa_J23114</partinfo>
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* <partinfo>BBa_J23116</partinfo>
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* <partinfo>BBa_J23118</partinfo>
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* <partinfo>pSB4C5</partinfo> - <partinfo>BBa_J23100</partinfo> (wiki name: BBa_J23100_4C5)
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* <partinfo>pSB4C5</partinfo> - <partinfo>BBa_J23101</partinfo> (wiki name: BBa_J23101_4C5)
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* <partinfo>pSB4C5</partinfo> - <partinfo>BBa_J23105</partinfo> (wiki name: BBa_J23105_4C5)
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* <partinfo>pSB4C5</partinfo> - <partinfo>BBa_J23106</partinfo> (wiki name: BBa_J23106_4C5)
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* <partinfo>pSB4C5</partinfo> - <partinfo>BBa_J23110</partinfo> (wiki name: BBa_J23110_4C5)
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* <partinfo>pSB4C5</partinfo> - <partinfo>BBa_J23114</partinfo> (wiki name: BBa_J23114_4C5)
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* <partinfo>pSB4C5</partinfo> - <partinfo>BBa_J23116</partinfo> (wiki name: BBa_J23116_4C5)
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* <partinfo>pSB4C5</partinfo> - <partinfo>BBa_J23118</partinfo> (wiki name: BBa_J23118_4C5)
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* <partinfo>BBa_B0031</partinfo> (negative control)
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==RESULTS==
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[[Image:UNIPV_Pavia_HCpromotersM9.png| 620px | center ]]
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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 promoters.
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{| border='1' align='center'
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|| Promoter || R.P.U. (average) || std
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|-
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|| <partinfo>BBa_J23100</partinfo> || 0.946 || 0.182
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|-
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|| <partinfo>BBa_J23101</partinfo> || 1 || -
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|-
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|| <partinfo>BBa_J23105</partinfo> || 0.196 || 0.038
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|-
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|| <partinfo>BBa_J23106</partinfo> || 0.326 || 0.015
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|-
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|| <partinfo>BBa_J23110</partinfo> || 0.678 || 0.096
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|-
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|| <partinfo>BBa_J23114</partinfo> || 0.091 || 0.022
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|-
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|| <partinfo>BBa_J23116</partinfo> || 0.101 || 0.004
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|-
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|| <partinfo>BBa_J23118</partinfo> || 0.655 || 0.035
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|}
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[[Image:UNIPV_Pavia_LCpromotersM9.png| 620px | center ]]
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In low copy number plasmids there are the same 3 groups but <partinfo>BBa_J23118</partinfo> resulted stronger than <partinfo>BBa_J23110</partinfo>.
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 +
{| border='1' align='center'
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|| Promoter || R.P.U. (average) || std
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|-
 +
|| <partinfo>pSB4C5</partinfo> - <partinfo>BBa_J23100</partinfo> || 0.838 || 0.101
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|-
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|| <partinfo>pSB4C5</partinfo> - <partinfo>BBa_J23101</partinfo> || 1 || -
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|-
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|| <partinfo>pSB4C5</partinfo> - <partinfo>BBa_J23105</partinfo> || 0.149 || 0.045
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|-
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|| <partinfo>pSB4C5</partinfo> - <partinfo>BBa_J23106</partinfo> || 0.286 || 0.033
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|-
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|| <partinfo>pSB4C5</partinfo> - <partinfo>BBa_J23110</partinfo> || 0.466 || 0.101
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|-
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|| <partinfo>pSB4C5</partinfo> - <partinfo>BBa_J23114</partinfo> || 0.053 || 0.048
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|-
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|| <partinfo>pSB4C5</partinfo> - <partinfo>BBa_J23116</partinfo> || 0.110 || 0.007
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|-
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|| <partinfo>pSB4C5</partinfo> - <partinfo>BBa_J23118</partinfo> || 0.714 || 0.154
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|}
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<td align="left" width="50%">[[Team:UNIPV-Pavia/Material Methods/Measurements/Tecan/test1settembre|Previous Test]]</td>
<td align="left" width="50%">[[Team:UNIPV-Pavia/Material Methods/Measurements/Tecan/test1settembre|Previous Test]]</td>
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<td align="right" width="50%">[[Team:UNIPV-Pavia/Material Methods/Measurements/Tecan/test14settembre| Next Test]]</td>
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<td align="right" width="50%">[[Team:UNIPV-Pavia/Material Methods/Measurements/Tecan/test8settembre| Next Test]]</td>
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{{UNIPV-Pavia/Menu_measurments}}
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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 14:10, 27 October 2010
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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 promoters.

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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