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 | 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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Revision as of 10:02, 15 April 2015
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04/14/2015
I ran a digest verification of Ryan's receiver [plasmid] with EcoRI and KpnI I also ran 3 ligations with: EsaI, RpaI, and RhiI with the gel-purified backbone and alkaline phosphatase treated inserts. | |
