Team:Purdue/Results/Wetlab Experiments

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<p>Our 2 main wet lab projects were the transformation of Bacillus Subtilis and assembly of our plasmids using Gibson Assembly.  
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<p>Our two main wet lab projects were the transformation of Bacillus Subtilis and assembly of our plasmids using Gibson Assembly.</p>
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We first conducted growth curve assays of our two strains (3A37 and 1A436) of Bacillus Subtilis. </p>
 
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'''Transformation of Bacillus Subtilis
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'''
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<p>We first conducted growth curve assays of our two strains (3A37 and 1A436) of Bacillus Subtilis obtained from The Ohio State Bacillus Stock Center . </p>
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<html><div z-index:100><img src="https://static.igem.org/mediawiki/2014/8/86/Phytosiddrophore_Assay2.PNG" width="600" height="600" align="right"></div></html>
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<p>Paper
 
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http://ac.els-cdn.com/S0167701207001315/1-s2.0-S0167701207001315-main.pdf?_tid=0f6ae27a-f585-11e3-a635-00000aacb361&acdnat=1402943939_6d4bbe673ecc5dfa11e7309507a2c4e8</p>
 
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'''Results'''
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<html><div z-index:100><img src="https://static.igem.org/mediawiki/2014/9/91/SAM_0252.jpg" width="550" height="450" align="center"></div></html>
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<p>We synthesized our 2 plasmids using Integrated DNA Technologies (IDT). We currently have them as gBlocks and are trouble shooting the Gibson Assembly.</p>
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Once we have functional circuits we will conduct the following Pytosidderophore Assay adapted from:
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Pérez-Miranda, S., Cabirol, N., George-Téllez, R., Zamudio-Rivera, L. S., & Fernández, F. J. (2007). O-CAS, a fast and universal method for siderophore detection. Journal of microbiological methods, 70(1), 127-131.
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<html><div z-index:100><img src="https://static.igem.org/mediawiki/2014/8/86/Phytosiddrophore_Assay2.PNG" width="550" height="550" align="center"></div></html>
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'''Gibson Assembly
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'''
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<p>We synthesized our two plasmids using Integrated DNA Technologies (IDT). We currently have them as gBlocks and are trouble shooting the Gibson Assembly.</p>

Latest revision as of 03:45, 18 October 2014

Wet Lab Experiments

Our two main wet lab projects were the transformation of Bacillus Subtilis and assembly of our plasmids using Gibson Assembly.


Transformation of Bacillus Subtilis

We first conducted growth curve assays of our two strains (3A37 and 1A436) of Bacillus Subtilis obtained from The Ohio State Bacillus Stock Center .


Results




Once we have functional circuits we will conduct the following Pytosidderophore Assay adapted from:

Pérez-Miranda, S., Cabirol, N., George-Téllez, R., Zamudio-Rivera, L. S., & Fernández, F. J. (2007). O-CAS, a fast and universal method for siderophore detection. Journal of microbiological methods, 70(1), 127-131.



Gibson Assembly

We synthesized our two plasmids using Integrated DNA Technologies (IDT). We currently have them as gBlocks and are trouble shooting the Gibson Assembly.