IGEM:Indian Institue of Technology Madras/2009/Notebook/PLASMID - Plasmid Locking Assembly for Sustaining Multiple Insert DNA/2009/09/28

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Entry title

 * OD v/s Conc expt: (DH5aplha; K12Z1; DH5aplha(P2 Plasmid); lacI- cells(P2 plasmid))
 * 3ml inoculum prepared.
 * 1.5ml added to Flasks
 * OD and conc were measured every 30 mins.
 * OD was calculated after resspending the cell pellet in Saline.
 * AHL confirmation:
 * AHL experiment: (DH5alpha; P2 in LacI- cells; T9005 cells)
 * Incubate all 3 cell types.
 * 2 mixtures made:
 * 1) 2ml T9005+ 2ml LacI- + 2 ml 2xyt broth -> with Amp
 * 2) 2ml T9005+ 2ml DH5aplha + 2 ml 2xyt broth --> without Amp
 * GFP expt:
 * inoculate DH5alpha cells in 5ml culture.
 * Add IPTG 3 hrs after incubation
 * Measure CFP after 5 hrs
 * The following was done as controls for restriction; ligation; Alkaline phosphatase treatment; and Transformation.
 * Restricted J23119(1A3) with S/P.
 * Restricted K156011(PCR) with X/P.
 * Alkaline Phosphatase (AP) of half of J23119(S/P) and half of K156011(X/P)
 * Following ligations performed: (AP = alkaline phosphatase treated)
 * C1) J23119(AP) + K156011;
 * C2) J23119(AP) + K156011(AP);
 * C3) J23119 + K156011(AP);
 * C4) J23119 + K156011;
 * C5) J23119(AP)
 * C6) J23119
 * Transformations of:
 * C1 through C6; C7(unligated J23119(S/P)); C8(unligated J23119(AP)(S/P))


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