Haynes Lab:Notebook/Characterizing AHL quorum sensing homologs/2015/04/14: Difference between revisions

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I ran a digest verification of Ryan's receiver [[https://benchling.com/s/VFNzuauP/edit|plasmid]] <br>  
I ran a digest verification of Ryan's receiver [[https://benchling.com/s/VFNzuauP/edit| plasmid]] with EcoRI and KpnI<br>  
Ryan cloned in an inducible promoter (digestion at BbsI site) and we want to check if it worked. <Br>  
Ryan cloned in an inducible promoter (digestion at BbsI site) and if it worked, it should remove the Kpnl cut site, resulting in one large band around ~3200 <Br>  
[[Image:14.4.15 Las inducible promoter gel image .tiff|x300px]]
[[Image:14.4.15 Las inducible promoter gel image .tiff|x300px]] <br>
The gel image shows two bands at around ~2200 and ~1000, which indicates that the inducible promoter did not clone in properly and the Kpnl cut site was still there. <br><Br>


I also ran 3 ligations with: EsaI, RpaI, and RhiI with the gel-purified backbone and alkaline phosphatase treated inserts. <br>
(3:1 ratio, 48ng insert per 50ng backbone). <br>


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Latest revision as of 00:54, 27 September 2017

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04/14/2015

I ran a digest verification of Ryan's receiver [plasmid] with EcoRI and KpnI
Ryan cloned in an inducible promoter (digestion at BbsI site) and if it worked, it should remove the Kpnl cut site, resulting in one large band around ~3200

The gel image shows two bands at around ~2200 and ~1000, which indicates that the inducible promoter did not clone in properly and the Kpnl cut site was still there.

I also ran 3 ligations with: EsaI, RpaI, and RhiI with the gel-purified backbone and alkaline phosphatase treated inserts.
(3:1 ratio, 48ng insert per 50ng backbone).


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