Week 3
From 2007.igem.org
(Difference between revisions)
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*We amplify some bio-bricks of interest to our project from the IGEM plates. We re-suspend and transform: | *We amplify some bio-bricks of interest to our project from the IGEM plates. We re-suspend 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_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_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_J04500 J04500], the PLac promoter inducible by IPTG; | ||
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::'''07/18/07''' | ::'''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 | + | *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 5ml of LB medium during the day. In the afternoon we dilute the 5 ml cultures in 50ml and let them grow overnight. |
| - | *We pick a colony from 07/17 plates, inoculate each in | + | *We pick a colony from 07/17 plates, inoculate each one in 5ml of LB medium and incubate overnight. |
::'''07/19/07''' | ::'''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 | + | *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 1mM IPTG to induce GFP expression to test fluorescence after 2h. ([[photos]]). |
| - | *Although [http://partsregistry.org/Part:BBa_J04431 J04431] works | + | *Although [http://partsregistry.org/Part:BBa_J04431 J04431] works correctly 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 right molecular weight. We think that maybe there's something wrong with the plasmid. |
Revision as of 09:20, 5 September 2007
- 07/17/07
- We amplify some bio-bricks of interest to our project from the IGEM plates. We re-suspend and transform:
- 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 J04430 and J04431 plates and grow it in 5ml of LB medium during the day. In the afternoon we dilute the 5 ml cultures in 50ml and let them grow overnight.
- We pick a colony from 07/17 plates, inoculate each one in 5ml of LB medium and incubate overnight.
- 07/19/07
- In the morning we grow the J04430 and J04431 cultures to an OD = 0.4 – 0.6. We add 1mM IPTG to induce GFP expression to test fluorescence after 2h. (photos).
- Although J04431 works correctly in presence of IPTG, J04430 doesn't. So, we perform a run on electrophoresis gel for the J04430 plasmid. We find that it doesn't match the right molecular weight. We think that maybe there's something wrong with the plasmid.
- 07/20/07
- Plasmid digestion (link):we digest J04500 with Spe1 and Pst1 and J04631 with Xba1 and Pst1.
- We perform the gel extraction procedure and store at -20°C.
- We amplify I13507 and R0051from the IGEM plates.
