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

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<h2>Kombucha Strains </h2>
 
<h2>Kombucha Strains </h2>
 
<p>The first steps in the characterization of microbes native to kombucha involved the isolation of strains from store-bought kombucha samples. This was accomplished by plating various dilutions of kombucha onto a variety of media including YPD, HS, and R2A.  </html>
 
<p>The first steps in the characterization of microbes native to kombucha involved the isolation of strains from store-bought kombucha samples. This was accomplished by plating various dilutions of kombucha onto a variety of media including YPD, HS, and R2A.  </html>
[[File:T--Austin_UTexas--IsolatingStrains.png|thumb|center|640px|Figure 1:Shows YPD plates spread with various dilutions of GT's brand kombucha samples. ]]
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[[File:T--Austin_UTexas--IsolatingStrains.png|thumb|center|640px|"'Figure 1:"' Shows YPD plates spread with various dilutions of GT's brand kombucha samples. ]]
 
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Isolated colonies were selected from each "isolation plate" and continually grown up and streaked out to ensure that the resulting frozen stock was truly axenic. Each newly isolated microbe was designated with a "KOM #" based on the order in which it was isolated (i.e. KOM 01, KOM 02, etc.) to serve as a placeholder name until the species could be identified. In order to begin this identification process, genomic DNA (gDNA) was first isolated from each individual strain. This DNA was then used as the template for two separate PCR reactions targeting either the 16S rRNA gene in bacteria, or the ITS rRNA gene for fungi. PCR products were then run on a 1% agarose gel to observe which reaction yielded product in gel.  
 
Isolated colonies were selected from each "isolation plate" and continually grown up and streaked out to ensure that the resulting frozen stock was truly axenic. Each newly isolated microbe was designated with a "KOM #" based on the order in which it was isolated (i.e. KOM 01, KOM 02, etc.) to serve as a placeholder name until the species could be identified. In order to begin this identification process, genomic DNA (gDNA) was first isolated from each individual strain. This DNA was then used as the template for two separate PCR reactions targeting either the 16S rRNA gene in bacteria, or the ITS rRNA gene for fungi. PCR products were then run on a 1% agarose gel to observe which reaction yielded product in gel.  
 
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<center><img src="https://static.igem.org/mediawiki/2016/4/44/T--Austin_UTexas--ExampleGel2.png" alt="RecapitulationsDay1" style="width:80%;"></center>
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[[File:T--Austin_UTexas--ExampleGel2.png|thumb|center|640px|"'Figure 2:"' Example of a gel obtained after running PCR products. Lane #1 is a 100 bp ladder; Lanes 2-7 are six different kombucha isolates that underwent PCR reactions selecting for bacteria (targeting 16S rRNA gene); Lanes 8-13 are the same six isolates, in the same order, but which underwent reactions selecting for fungi (targeting ITS rRNA gene). Based on the gel results it can be observed that for any given isolate, a product in gel was only observed for either bacterial or fungal primer reactions.  ]]
<figcaption><b>Figure 2:</b>Example of a gel obtained after running PCR products. Lane #1 is a 100 bp ladder; Lanes 2-7 are six different kombucha isolates that underwent PCR reactions selecting for bacteria (targeting 16S rRNA gene); Lanes 8-13 are the same six isolates, in the same order, but which underwent reactions selecting for fungi (targeting ITS rRNA gene). Based on the gel results it can be observed that for any given isolate, a product in gel was only observed for either bacterial or fungal primer reactions.   
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Revision as of 17:40, 19 October 2016

Results


Click on one of the images below to learn more about our results!