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'''
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*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 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 5 ml of LB medium and incubate overnight.   
+
*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 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 1mM IPTG to induce GFP expression to test fluorescence after 2h. ([[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.
+
*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:
    • [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 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 [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 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.


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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