Week 3
From 2007.igem.org
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::'''07/17/07''' | ::'''07/17/07''' | ||
| - | *We amplify some bio- bricks of interest for our project from the IGEM plates. We | + | *We amplify some bio- bricks of interest for 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; | ||
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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 5 ml of LB medium during the day. In the afternoon we | + | *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 dilute 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. | *We pick a colony from 07/17 plates, inoculate each in 5 ml of LB medium and incubate overnight. | ||
| Line 19: | Line 19: | ||
*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]]). | *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]]). | ||
| - | * | + | *Although [http://partsregistry.org/Part:BBa_J04431 J04431] works right 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 maybe there's something wrong with the plasmid. |
Revision as of 09:10, 5 September 2007
- 07/17/07
- We amplify some bio- bricks of interest for 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_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 dilute 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).
- Although [http://partsregistry.org/Part:BBa_J04431 J04431] works right 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 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.
