Team:Glasgow/Results/PromoterLibrary/Results

From 2011.igem.org

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{{Team:Glasgow/Header}}
<h3>Results:</h3>
<h3>Results:</h3>
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<html>
<b><u>Stock of Promoters and Ribosome Binding Sites</b></u>
<b><u>Stock of Promoters and Ribosome Binding Sites</b></u>
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We have successfully created a stock of all the promoters and ribosome binding sites which we aimed to make.
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We have successfully created a stock of all the promoters and ribosome binding sites ligated to each other.
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<br/><br/>
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<br/><center><table border="1">
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<b><u>Stocks of Ligated Constructs</b></u>
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<tr>
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<br/><br/>
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<td>Strong Promoter + Strong RBS</td>
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We have successfully obtained colonies from the following ligations:
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<td>Strong Promoter + Weak RBS</td>
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</tr>
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Strong Promoter + Strong RBS<Br/>
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<tr>
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Strong Promoter + Weak RBS<br/>
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<td>Weak Promoter + Strong RBS</td>
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Weak Promoter + Strong RBS<br/>
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<td>Weak Promoter + Weak RBS</td>
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Weak Promoter + Weak RBS<br/>
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</tr>
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pBAD Promoter + Strong RBS<br/>
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<tr>
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pBAD promoter + Weak RBS<br/>
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<td>pBAD promoter + Strong RBS</td>
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Bluf Promoter + Strong RBS<br/>
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<td>pBAD promoter + Weak RBS</td>
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Bluf Promoter + Weak RBS<br/>
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</tr>
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OmpF Promoter + Strong RBS<br/>
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<tr>
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OmpF Promoter + Weak RBS<br/>
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<td>Bluf promoter + Strong RBS</td>
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OmpC Promoter + Strong RBS<br/>
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<td>Bluf promoter + Weak RBS</td>
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OmpC Promoter + Weak RBS <br/><br/>
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</tr>
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<td>OmpF promoter + Strong RBS</td>
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<td>OmpF promoter + Weak RBS</td>
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</tr>
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<tr>
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<td>OmpC promoter + Strong RBS</td>
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<td>OmpC promoter + Weak RBS</td>
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</table> </center><br/>As these constructs were made using AlwnI and gel extraction, any colony that successfully grew must have worked. However these parts were not present on the submission plasmid (pSB1C3). 
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<br/>These parts are all present in the plasmid which the promoter part was present in on the Registry.<br/>Upon ligating the parts into the submission vector (pSB1C3), only the following constructs successfully grew and were confirmed to be the right size on a gel:<br/>
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<br/>The whole fragment was digested out of the plasmid using EcoRI and PstI, and ligated into the linear submission vector.  At this stage only the following constructs successfully grew and were confirmed to be the right size on a gel:<br/><br/>
Bluf Promoter + Strong RBS<br/>
Bluf Promoter + Strong RBS<br/>
Bluf Promoter + Weak RBS<br/>
Bluf Promoter + Weak RBS<br/>
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Weak Promoter + Weak RBS<br/><br/>
Weak Promoter + Weak RBS<br/><br/>
<center>
<center>
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<table border="1">
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<tr>
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<img src=https://static.igem.org/mediawiki/2011/3/39/Glasgow_gel_prom_all.png>
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<td><img src=https://static.igem.org/mediawiki/2011/1/1d/Glasgowgel_-_bluf_%2B_prom.png width=250 height=250></td>
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<br/><i>Gels 1 to 4 show the Promoter + RBS constructs we made that were ligated into the submission vector and found to be the expected size</i><Br/>
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<td><img src=https://static.igem.org/mediawiki/2011/3/39/Glasgowgel_-_weak_prom_%2B_s.rbs.png width=250 height=250></td>
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</tr>
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<tr>
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<td><img src=https://static.igem.org/mediawiki/2011/d/db/Glasgowgel_-_w.prom_%2B_w.rbs.png width=250 height=250></td>
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<td><img src=https://static.igem.org/mediawiki/2011/4/4f/Glasgowgel_-_ompf.png width=250 height=250></td>
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</tr>
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</table>
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Latest revision as of 03:06, 22 September 2011

Results:

Stock of Promoters and Ribosome Binding Sites
We have successfully created a stock of all the promoters and ribosome binding sites ligated to each other.

Strong Promoter + Strong RBS Strong Promoter + Weak RBS
Weak Promoter + Strong RBS Weak Promoter + Weak RBS
pBAD promoter + Strong RBS pBAD promoter + Weak RBS
Bluf promoter + Strong RBS Bluf promoter + Weak RBS
OmpF promoter + Strong RBS OmpF promoter + Weak RBS
OmpC promoter + Strong RBS OmpC promoter + Weak RBS

As these constructs were made using AlwnI and gel extraction, any colony that successfully grew must have worked. However these parts were not present on the submission plasmid (pSB1C3).

The whole fragment was digested out of the plasmid using EcoRI and PstI, and ligated into the linear submission vector. At this stage only the following constructs successfully grew and were confirmed to be the right size on a gel:

Bluf Promoter + Strong RBS
Bluf Promoter + Weak RBS
OmpF + Weak RBS
pBAD + Strong RBS
Weak Promoter + Strong RBS
Strong Promoter + Strong RBS
Weak Promoter + Weak RBS


Gels 1 to 4 show the Promoter + RBS constructs we made that were ligated into the submission vector and found to be the expected size

Retrieved from "http://2011.igem.org/Team:Glasgow/Results/PromoterLibrary/Results"