Week 3

From 2007.igem.org

Revision as of 10:18, 28 August 2007

07/17/07

• We amplified some bio- bricks of interest for our project from the IGEM plates. We resuspended and transformed:
  • [http://partsregistry.org/Part:BBa_J04430 J04430] to test the GFP fluorescence detection with our experimental set up;
  • [http://partsregistry.org/Part:BBa_J04431 J04431] to test the GFP (+LVA) half-life since we needed a GFP with a short one for our project;
  • [http://partsregistry.org/Part:BBa_J04500 J04500], the PLac promoter inducible by IPTG;
  • [http://partsregistry.org/Part:BBa_J04631 J04631], the GFP (+LVA) protein.
• We straked on plates with the right antibiotic.

07/18/07

• 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.

07/19/07

• 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. (link results foto).
• 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.

07/20/07

• 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 R0051 from the IGEM plate.

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