Week 3
From 2007.igem.org
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*''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. | *''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 perform the gel extraction procedure and store at -20°C. | ||
| - | *We amplify | + | *We amplify [http://partsregistry.org/Part:BBa_I13507 I13507] and [http://partsregistry.org/Part:BBa_R0051 R0051]from the IGEM plates. |
[[Bologna | Back]] | [[Bologna | Back]] | ||
Revision as of 15:11, 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 electroforesis 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.
