Week 3

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Revision as of 12:18, 28 August 2007 by Francesca.bugane2 (Talk | contribs)
  • We amplified some bio- bricks of interest for our project from the IGEM plates. We resuspended and transformed:
    • J04430 to test the GFP fluorescence detection with our experimental set up;
    • J04431 to test the GFP (+LVA) half-life since we needed a GFP with a short one for our project;
    • J04500, the PLac promoter inducible by IPTG;
    • J04631, the GFP (+LVA) protein.
  • We straked on plates with the right antibiotic.

  • We performed a test for GFP induction with IPTG. We picked a colony from the J04430 and J04431 plates and grew it in 5 ml of LB medium during the day. In the afternoon we diluited the 5 ml cultures in 50 ml and let them growing overnight.
  • We picked a colony from 07/17 plates and inoculated in 5 ml of LB medium from each plate and incubated overnight.

  • In the morning we diluited the J04430 and J04431 cultures to an OD=0.05. We grew the cultures till an OD = 0.4 – 0.6. We added 1 mM IPTG to induce GFP expression and we tested fluorescence after 2 h. (photos).
  • Althought J04431 worked rigth in presence of IPTG, J04430 didn't. So, we performed a run on electroforesis gel for the J04430 plasmid. We found that it didn't match the rigth molecular weight. We thougth maybe there's something wrong with the plasmid.

  • Plasmid digestion (link):we digested J04500 with Spe1 and Pst1 and J04631 with Xba1 and Pst1.
  • We performed the gel extraction procedure and store at -20°C.
  • We amplified amd transformed I13507 and R0051from the IGEM plate.