Week 3

From 2007.igem.org

Revision as of 10:27, 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 [http://partsregistry.org/Part:BBa_J04430 J04430] and [http://partsregistry.org/Part:BBa_J04431 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 [http://partsregistry.org/Part:BBa_J04430 J04430] and [http://partsregistry.org/Part:BBa_J04431 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 [http://partsregistry.org/Part:BBa_J04431 J04431] worked rigth in presence of IPTG, [http://partsregistry.org/Part:BBa_J04430 J04430] didn't. So, we performed a run on electroforesis gel for the [http://partsregistry.org/Part:BBa_J04430 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 [http://partsregistry.org/Part:BBa_J04500 J04500] with Spe1 and Pst1 and [http://partsregistry.org/Part:BBa_J04631 J04631] with Xba1 and Pst1.
• We performed the gel extraction procedure and store at -20°C.
• We amplified amd transformed [http://partsregistry.org/Part:BBa_I13507 I13507] and [http://partsregistry.org/Part:BBa_R0051 R0051]from the IGEM plate.

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