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

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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
File:14.4.15 Las inducible promoter gel image .tiff
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).