Week 3

From 2007.igem.org

Revision as of 15:28, 3 September 2007

07/17/07

• We amplify some bio- bricks of interest for our project from the IGEM plates. We resuspend and transform:
  • [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 need 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 strake on plates with the right antibiotic.

07/18/07

• We perform a test for GFP induction with IPTG. We pick a colony from the [http://partsregistry.org/Part:BBa_J04430 J04430] and [http://partsregistry.org/Part:BBa_J04431 J04431] plates and grow it in 5 ml of LB medium during the day. In the afternoon we diluite the 5 ml cultures in 50 ml and let them growing overnight.

• We pick a colony from 07/17 plates, inoculate each in 5 ml of LB medium and incubate overnight.

07/19/07

• In the morning we grow the [http://partsregistry.org/Part:BBa_J04430 J04430] and [http://partsregistry.org/Part:BBa_J04431 J04431] cultures to an OD = 0.4 – 0.6. We add 1 mM IPTG to induce GFP expression to test fluorescence after 2 h. (photos).
• Althought [http://partsregistry.org/Part:BBa_J04431 J04431] works rigth in presence of IPTG, [http://partsregistry.org/Part:BBa_J04430 J04430] doesn't. So, we perform a run on electrophoresis gel for the [http://partsregistry.org/Part:BBa_J04430 J04430] plasmid. We find that it doesn't match the rigth molecular weight. We think maybe there's something wrong with the plasmid.

07/20/07

• Plasmid digestion (link):we digest [http://partsregistry.org/Part:BBa_J04500 J04500] with Spe1 and Pst1 and [http://partsregistry.org/Part:BBa_J04631 J04631] with Xba1 and Pst1.
• We perform the gel extraction procedure and store at -20°C.
• We amplify [http://partsregistry.org/Part:BBa_I13507 I13507] and [http://partsregistry.org/Part:BBa_R0051 R0051]from the IGEM plates.

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