Week 3
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Revision as of 13:25, 20 August 2007 by Alice.pasini (Talk | contribs)
- 07/17/07
- We amplified some bio- bricks of interest for our project from the IGEM plates. We resuspended and transformed:
-BBa_ J04430 to test the GFP fluorescence detection with our experimental set up
-BBa_J04431 to test the GFP (+LVA) half-life since we needed a GFP with a short one for our project.
- BBa_J04500, the PLac promoter inducible by IPTG;
- BBa_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 plate and grew it till a OD 0.4 – 0.6. We added 1 mM IPTG to induce GFP expression.
- 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
- We induced GFP expression from J04431 and J04430. Althought the first worked rigth in presence of IPTG, the second 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.