IGEM:Paris Bettencourt 2012/Notebooks/suicide group/day by day//2012/09/08

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(Colicin Assay)
(Colicin Assay)
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Given the puzzling results of the standard Colicin E2 assay (sparse lawns, apparent loss of RFP plasmid), we surmised that the lawn bacteria were at some point contaminated with Colicin E2. We therefore selected 3 colonies from the purportedly contaminated assays to spot on WT(MG1655) lawns. This seems to be a satisfying hypothesis only for the NEB.08.23 cells, but not the Z1.08.23 cells.
Given the puzzling results of the standard Colicin E2 assay (sparse lawns, apparent loss of RFP plasmid), we surmised that the lawn bacteria were at some point contaminated with Colicin E2. We therefore selected 3 colonies from the purportedly contaminated assays to spot on WT(MG1655) lawns. This seems to be a satisfying hypothesis only for the NEB.08.23 cells, but not the Z1.08.23 cells.
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[[Image:SGNEB0823C2060912.jpg |350px |thumb|ColE2 spotted on NEB.08.23 (NEB+ColE2 IMM+RFP) cell lawn]]
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{|
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[[Image:SGZ10823C2060912.jpg |350px |thumb|ColE2 spotted on Z1.08.23 (Z1+ColE2 IMM+RFP) cell lawn]]
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| [[Image:SGNEB0823C2060912.jpg |350px |thumb|ColE2 spotted on NEB.08.23 (NEB+ColE2 IMM+RFP) cell lawn]]
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| [[Image:SGZ10823C2060912.jpg |350px |thumb|ColE2 spotted on Z1.08.23 (Z1+ColE2 IMM+RFP) cell lawn]]
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|}
Colonies (1,2,3) were selected from each plate to spot on WT lawns to determine if these colonies were in fact Colicin E2 cells.  
Colonies (1,2,3) were selected from each plate to spot on WT lawns to determine if these colonies were in fact Colicin E2 cells.  

Revision as of 06:32, 17 September 2012

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Colicin Assay

Given the puzzling results of the standard Colicin E2 assay (sparse lawns, apparent loss of RFP plasmid), we surmised that the lawn bacteria were at some point contaminated with Colicin E2. We therefore selected 3 colonies from the purportedly contaminated assays to spot on WT(MG1655) lawns. This seems to be a satisfying hypothesis only for the NEB.08.23 cells, but not the Z1.08.23 cells.

ColE2 spotted on NEB.08.23 (NEB+ColE2 IMM+RFP) cell lawn
ColE2 spotted on NEB.08.23 (NEB+ColE2 IMM+RFP) cell lawn
ColE2 spotted on Z1.08.23 (Z1+ColE2 IMM+RFP) cell lawn
ColE2 spotted on Z1.08.23 (Z1+ColE2 IMM+RFP) cell lawn

Colonies (1,2,3) were selected from each plate to spot on WT lawns to determine if these colonies were in fact Colicin E2 cells.

Positive Control, ColE2 spotted on Z1.23 (Z1+RFP) cell lawn
Positive Control, ColE2 spotted on Z1.23 (Z1+RFP) cell lawn
Negative Control, MG1655 spotted on Z1.23 (Z1+RFP) cell lawn
Negative Control, MG1655 spotted on Z1.23 (Z1+RFP) cell lawn
Positive Control, ColE2 spotted on MG1655 cell lawn
Positive Control, ColE2 spotted on MG1655 cell lawn
NEB.08.23 Colony 1 spotted on MG1655 cell lawn
NEB.08.23 Colony 1 spotted on MG1655 cell lawn
NEB.08.23 Colony 2 spotted on MG1655 cell lawn
NEB.08.23 Colony 2 spotted on MG1655 cell lawn
NEB.08.23 Colony 3 spotted on MG1655 cell lawn
NEB.08.23 Colony 3 spotted on MG1655 cell lawn
Z1.08.23 Colony 1 spotted on MG1655 cell lawn
Z1.08.23 Colony 1 spotted on MG1655 cell lawn
Z1.08.23 Colony 2 spotted on MG1655 cell lawn
Z1.08.23 Colony 2 spotted on MG1655 cell lawn
Z1.08.23 Colony 3 spotted on MG1655 cell lawn
Z1.08.23 Colony 3 spotted on MG1655 cell lawn


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