IGEM:Virginia/2012/Notebook/Genetically engineered bacteriophage for diagnosis of whooping cough/2012/09/16

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(Autocreate 2012/09/16 Entry for IGEM:Virginia/2012/Notebook/Genetically_engineered_bacteriophage_for_diagnosis_of_whooping_cough)
Current revision (20:07, 3 October 2012) (view source)
 
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|style="background-color: #EEE"|[[Image:igem-logo-150px.png|150px]]<span style="font-size:22px;"> iGEM Project name 1</span>
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|style="background-color: #EEE"|[[Image:igem-logo-150px.png|150px]]<span style="font-size:22px;">Week of 9/16/12</span>
|style="background-color: #F2F2F2" align="center"|<html><img src="/images/9/94/Report.png" border="0" /></html> [[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>[[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]]<html>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]]<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>}}
|style="background-color: #F2F2F2" align="center"|<html><img src="/images/9/94/Report.png" border="0" /></html> [[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>[[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]]<html>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]]<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>}}
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==Entry title==
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==Week of 9/16/12==
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* Insert your content here.
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Performed gel extraction of ligated backbones (both E+P and E+S); failed to show up in
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adequate amounts when nanodropped.
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<br>
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Ligated Plasmid+RBS (E+S), SynCG (X+P), and RFP (X+P) at 3:1 and 1:1 ratios of both
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2X ligase buffer and promega DNA ligase and performed PCR.
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<br>
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Repeated ligation and performed PCR purification: P+RBS+RFP and P+RBS+SynCG
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demonstrate proper amounts of DNA but at low concentrations, suggesting that perhaps
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the DNA is not the intended amplicon.
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<br>
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Discovered that roughly half of the previous pregnancy tests now appear positive,
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brighter than evaporation lines would be. Some tests for 5 mM DTT (denaturing
 +
solution), 0.5 mM DTT, 0.05 mM DTT, and 0 mM DTT are now positive with the
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majority in the latter two groups.
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<br>
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Nanodropped the P+RBS+RFP, P+RBS+SynCG, backbone (E+S), backbone (E+P),
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and SynCG all (E+P): only the backbone (E+S) and SynCG all (E+P) showing proper
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concentrations. The latter includes the IDT plasmid backbone.
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<br>
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Plated the bbp9 biobricks (both 3:1 and 1:1 ratios), the hCG biobrick (in 3:1, 1:1, and
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1:3 ratios), backbone, original BBa_K331022, and a DNA lacking group, producing
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thousands of cells on each plate (except for the last), demonstrating that many of the cells
 +
have empty vectors.

Current revision

Week of 9/16/12 Main project page
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Week of 9/16/12

Performed gel extraction of ligated backbones (both E+P and E+S); failed to show up in adequate amounts when nanodropped.
Ligated Plasmid+RBS (E+S), SynCG (X+P), and RFP (X+P) at 3:1 and 1:1 ratios of both 2X ligase buffer and promega DNA ligase and performed PCR.
Repeated ligation and performed PCR purification: P+RBS+RFP and P+RBS+SynCG demonstrate proper amounts of DNA but at low concentrations, suggesting that perhaps the DNA is not the intended amplicon.
Discovered that roughly half of the previous pregnancy tests now appear positive, brighter than evaporation lines would be. Some tests for 5 mM DTT (denaturing solution), 0.5 mM DTT, 0.05 mM DTT, and 0 mM DTT are now positive with the majority in the latter two groups.
Nanodropped the P+RBS+RFP, P+RBS+SynCG, backbone (E+S), backbone (E+P), and SynCG all (E+P): only the backbone (E+S) and SynCG all (E+P) showing proper concentrations. The latter includes the IDT plasmid backbone.
Plated the bbp9 biobricks (both 3:1 and 1:1 ratios), the hCG biobrick (in 3:1, 1:1, and 1:3 ratios), backbone, original BBa_K331022, and a DNA lacking group, producing thousands of cells on each plate (except for the last), demonstrating that many of the cells have empty vectors.



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