Jacqueline K. Grimm at 00:09, 4 October 2012

2012-10-04T00:09:08Z

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-	\|style="background-color: #EEE"\|[[Image:igem-logo-150px.png\|150px]]<span style="font-size:22px;"> ~~iGEM Project name 1~~</span>	+	\|style="background-color: #EEE"\|[[Image:igem-logo-150px.png\|150px]]<span style="font-size:22px;"> Week of 9/23/12</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>      </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>      </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~~==	+	==Week of 9/23/12==
-	* Insert your content here.	+	Redid backbone (with both E+S and E+P) but CIAPing the vector prior to ligation
		+	in order to decrease the chance of the empty vector ligating to itself and creating
		+	false positive colonies, producing acceptable concentrations of 90.4 and 71.0 ng/μL,
		+	respectively. Colonies for the bbp9, hCG, SynCG+promotor, SynCG+promotor+RBS,,
		+	plasmid (E+S), and plasmid (E+P) all are acceptable in the double digits. The SynCG
		+	colony is still in the thousands. Inoculated colonies, performed plasmid prep, and
		+	used PCR to confirm presence of inserts. hCG, SynCG, and P+RBS+SynCG all have
		+	exceptionally large yields over 5000 ng/μL.
		+	<br>
		+	EtBr gel confirms presence of hCG biobrick and SynCG biobrick.
		+	<br>
		+	Extracted SynCG (E+X) with phenolchloroform, resulting in similarly high yields.
		+	<br>
		+	Attempted ligations of bbp9 biobrick but only empty vectors were produced.
		+	<br>
		+	Transformed DH5α with ligations and plated on LB with chlor and made inoculations
		+	of bbp9 colonies and incubated at 37°C. The colonies later grew and were turned into 5
		+	5mL cultures.
		+	<br>
		+	Plasmid prep for 5 bbp9 inoculations and 5 P+RBS+SynCG inoculation and then run
		+	PCR and gel. All colonies were acceptable with a concentration of at least 3000 ng/μL.
		+	<br>
		+	Gel indicates that the fourth bbp9 PCR may have a possible biobrick as well as the first
		+	P+RBS+SynCG.
		+	<br>
		+	Restarted with 6 tube of T7 digest with BclI, followed by ethanol precipitation cleanup
		+	and a PCR cleanup kit. Cleaned up the T7 halves and resuspended them in 20 μL of EB
		+	buffer, resulting in a final concentration of 623,1 ng/μL.
		+	<br>
		+	Performed PCR amplification of promotor, RBS, SynCG after plasmid prep of DH5α
		+	transformed with biobrick plasmid. No evidence of primers appeared after running the
		+	PCR products on a 2% agarose gel, suggesting that the PCR mixture was incorrect.
		+	<br>
		+	Performed 12 inoculations of P+RBS+SynCG colonies. After running these colonies
		+	through a PCR, the nanodropping of these plasmid preps confirmed that all 12 had
		+	acceptable concentrations in the upper double digits.

Jacqueline K. Grimm: Autocreate 2012/09/23 Entry for IGEM:Virginia/2012/Notebook/Genetically_engineered_bacteriophage_for_diagnosis_of_whooping_cough

2012-10-04T00:07:38Z

Autocreate 2012/09/23 Entry for IGEM:Virginia/2012/Notebook/Genetically_engineered_bacteriophage_for_diagnosis_of_whooping_cough

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|style="background-color: #EEE"|[[Image:igem-logo-150px.png|150px]]<span style="font-size:22px;"> iGEM Project name 1</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>      </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==
* Insert your content here.


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__NOTOC__

IGEM:Virginia/2012/Notebook/Genetically engineered bacteriophage for diagnosis of whooping cough/2012/09/23 - Revision history

Jacqueline K. Grimm at 00:09, 4 October 2012

Jacqueline K. Grimm: Autocreate 2012/09/23 Entry for IGEM:Virginia/2012/Notebook/Genetically_engineered_bacteriophage_for_diagnosis_of_whooping_cough