User:Karmella Haynes/Notebook/BioBrick cloning/2011/05/31

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05/31/11

 * &#x2713; Assemblies: Re-build mIR sensor test constructs without insulators and with CFP-Reporter, no PEST

Assemblies --> Old plasmids are very problematic. As long as I'm rebuilding, may as well update the initial miR sensor test experiments to match the current design --> Build complete "matrix" of conditions: --> Differences from previous constructs: No HS4 insulators, final constructs in MV8, no PEST on CFP, polyA2 (Mara's M127)
 * Px (RFP-repressor promoter) = Ubc, HPK, CMV
 * Py (CFP-reporter promoter) = Ubc, HPK, CMV
 * Gal4 binding sites = 0, 5x, 10x


 * 1) KAH185: KAH83/(E/S)/443 &#x2713; + 5xGal4/(E/X)/~3293 &#x2713;
 * 2) KAH186: Ubc/(E/S)/344 &#x2713; + (1) KAH182/(E/X)/~4875
 * 3) KAH187: CMV/(E/S)/588 &#x2713; + "
 * 4) KAH188: KAH83/(E/S)/443 + "
 * 5) KAH189: KAH84/(E/S)/615 + "
 * 6) KAH190: KAH85/(E/S)/687 + "
 * 7) KAH192: (2) FC6/(E/S)/780 + "
 * 8) KAH193: Ubc/(E/S)/344 &#x2713; + KAH73/(E/X)/~5638 &#x2713;
 * 9) KAH194: CMV/(E/S)/588 &#x2713; + "

> Digests (Fermentas FD) --> Specific notes

> Measure conc.'s

> Ligations


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