Difference between revisions of "Team:Austin UTexas/Experiments"

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<p>Describe the experiments, research and protocols you used in your iGEM project.</p>-->
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Ongoing Experiments
 
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*Recapitulation
 
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*Sequencing
 
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*Conjugation
 
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*Ethanol Screening
 
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*pH Detection
 
  
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<h2>Ongoing Experiments</h2>
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<h3>Sequencing</h3>
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<p>By plating store-bought kombuchas onto a variety of media plates, we were able to isolate several microbial colonies. These colonies were harvested and made into frozen stocks for future use. Fresh cultures were innoculated with these isolated strains and allowed to grow until turbid. Using a genomic DNA (gDNA) isolation kit (Invitrogen), DNA was extracted from these unknown microbes. Two PCR reactions were performed for each microbe using the appropriate isolated gDNA as a template and bacterial primers (selects for bacteria and targets the 16s rRNA gene) or fungal primers (selects for fungi and targets ITS rRNA gene). The purpose of these reactions was to determine whether each isolated species was bacterial or fungal. The PCR products were run on a 1% agarose gel and those that yielded products in the gel were PCR purified using BioBasic and sent to ?????????????????? for sequencing (using Sanger sequencing). The resulting sequences were identified using the <a href = "https://rdp.cme.msu.edu/">Ribosomal Database Project.</a></p>
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<h3>Conjugation</h3>
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<p></p>
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<h3>Ethanol Screening</h3>
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<p></p>
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<h3>Recapitulation</h3>
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<p></p>
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<h3>pH Detection</h3>
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<p></p>
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<br>
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<br>
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<h2>Looking Forward</h2>
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<h3>Brazzein</h3>
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Upcoming
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*Brazzein
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*Leptin
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*G. Oxydans pH Detection
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Revision as of 23:33, 16 October 2016

Ongoing Experiments

Sequencing

By plating store-bought kombuchas onto a variety of media plates, we were able to isolate several microbial colonies. These colonies were harvested and made into frozen stocks for future use. Fresh cultures were innoculated with these isolated strains and allowed to grow until turbid. Using a genomic DNA (gDNA) isolation kit (Invitrogen), DNA was extracted from these unknown microbes. Two PCR reactions were performed for each microbe using the appropriate isolated gDNA as a template and bacterial primers (selects for bacteria and targets the 16s rRNA gene) or fungal primers (selects for fungi and targets ITS rRNA gene). The purpose of these reactions was to determine whether each isolated species was bacterial or fungal. The PCR products were run on a 1% agarose gel and those that yielded products in the gel were PCR purified using BioBasic and sent to ?????????????????? for sequencing (using Sanger sequencing). The resulting sequences were identified using the Ribosomal Database Project.

Conjugation

Ethanol Screening

Recapitulation

pH Detection



Looking Forward

Brazzein