Team:UC Chile/Protocols

From 2012.igem.org

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[[File: Chile_pipeting.jpg| 300px| right]]
[[File: Chile_pipeting.jpg| 300px| right]]
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<p>Here you will find various protocols we have used during the development of our project. We have separated them into 4 main sections: <br /><br />  [[#General Protocols]] <br /> [[#Cyanobacteria Protocols]] <br /> [[#Bactomithril Protocols]] <br /> [[#Characterization Protocols]] </p>
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<p>Here you will find various protocols we have used during the development of our project. We have separated them into 3 main sections: <br /><br />  [[#General Protocols]] <br /> [[#Cyanobacteria Protocols]] <br /> [[#Characterization Protocols]] </p>
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<div id="General Protocols">
<div id="General Protocols">
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Concentrate the DNA to be transformed into Synechocystis to 1µg/µL in 10µL for each transformation (A total of 10 ug of the transformation plasmid).  
Concentrate the DNA to be transformed into Synechocystis to 1µg/µL in 10µL for each transformation (A total of 10 ug of the transformation plasmid).  
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Inoculate Synechocystis PCC 6803 into a sterile 250mL Erlenmeyer flask with 150mL* of liquid BG-11 and grow culture until you reach an OD730nm of 0.8 1.0 (That should take between 6 to 10 days dependending on the amount of initial inoculum).  
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Inoculate Synechocystis PCC 6803 into a sterile 250mL Erlenmeyer flask with 150mL* of liquid BG-11 and grow culture until you reach an OD730nm of 0.4 0.6, never above 0.8 (That should take between 6 to 10 days dependending on the amount of initial inoculum).  
* For each 50 mL of Synechocystis PCC 6803 you will have 5 individual transformations.  
* For each 50 mL of Synechocystis PCC 6803 you will have 5 individual transformations.  
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<div id="Reinoculation">
<div id="Reinoculation">
<h2>Reinoculation</h2>
<h2>Reinoculation</h2>
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Protocol right here!
 
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</div>
 
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<div id="Cyanobacterial DNA extraction">
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<h2>Cyanobacterial DNA extraction</h2>
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This is a standard protocol for propagating cultures. The amount of initial innoculum will depend on when you will need a specific OD. Refer to [https://2012.igem.org/Team:UC_Chile/Results/Growthcurve growth curves] to calculate your needed inoculum through a standard C*V = constant relation.
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</div>
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<p>Protocol goes here!</p>
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<div id="Bactomithril Protocol">
 
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<h1>Bactomithril Protocols</h1>
 
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</div>
 
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<p>You can find a complete protocol to produce spider-silk fibers in the following paper:
 
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Teulé, F., Cooper, A. R., Furin, W. a, Bittencourt, D., Rech, E. L., Brooks, A., & Lewis, R. V. (2009). A protocol for the production of recombinant spider silk-like proteins for artificial fiber spinning. Nature protocols, 4(3), 341-55. doi:10.1038/nprot.2008.250
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All work must be done under sterile conditions.
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<br><br>Other easier protocols are in the following adresses:<br>
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http://www.pnas.org/content/107/32/14059.long<br>
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<li>Autoclave a flask with enough volume to harbor at least twice the volume you require</li>
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http://onlinelibrary.wiley.com/doi/10.1002/jbm.a.34353/full</p>
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<li>Measure and add BG-11 media to flask</li>
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<li>Add axenic Synechocystis to flask. Inoculum must be in range 5-20% v/v of final volume</li>
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</ul>
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</div>
<div id="Characterization Protocols">
<div id="Characterization Protocols">
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<div id="SyneGrowth">
<div id="SyneGrowth">
<h2>Methods for the characterization of Synechocystis PCC 6803 growth curve</h2>
<h2>Methods for the characterization of Synechocystis PCC 6803 growth curve</h2>
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This are the methods we used for setting up the growth curve experiment of our Synechocystis PCC 6803 which is further described [[Team:UC_Chile2/Characterization | here ]].
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These are the methods we used for setting up the growth curve experiment of our Synechocystis PCC 6803 which is further described [[Team:UC_Chile2/Characterization | here ]].
<h3>Materials</h3>
<h3>Materials</h3>

Latest revision as of 08:36, 26 October 2012

Project: Luxilla - Pontificia Universidad Católica de Chile, iGEM 2012