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

From 2010.igem.org

SEPTEMBER, 2ND

PLATE

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

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

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

Tecan Test