Difference between revisions of "IGEM:Cambridge/2008/Notebook/Turing Pattern Formation/2008/08/14"

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(BioBrick extraction for testing promoters and RBS in B.subtillis)
(BioBrick extraction for testing promoters and RBS in B.subtillis)
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We attempted to isolate 4 BioBricks: I13522 (GFP under constitutive promoter), E0040 (GFP only), R0040 (a promoter), & B0034 (an ''E.coli'' RBS).
 
We attempted to isolate 4 BioBricks: I13522 (GFP under constitutive promoter), E0040 (GFP only), R0040 (a promoter), & B0034 (an ''E.coli'' RBS).
So far, only the former two, extracted from 2007 wells, grew after transformation. Single-colony PCR was used to test plasmids.
+
So far, only the former two, extracted from 2007 wells, grew after transformation. Single-colony PCR was used to test the transformants.
  
 
Expected VF2-VR fragment sizes:
 
Expected VF2-VR fragment sizes:

Revision as of 16:11, 14 August 2008



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BioBrick extraction for testing promoters and RBS in B.subtillis

The primers for inducible B.subtillis promoters have been ordered. Meanwhile, we would like to be able to compare the RBS and promoter strengths in E.coli and B.subtillis, using GFP fluorescence to quantify gene expression.

We attempted to isolate 4 BioBricks: I13522 (GFP under constitutive promoter), E0040 (GFP only), R0040 (a promoter), & B0034 (an E.coli RBS). So far, only the former two, extracted from 2007 wells, grew after transformation. Single-colony PCR was used to test the transformants.

Expected VF2-VR fragment sizes:

I13522 - ~2.4 kb E0040 - 958 bp R0040 - 292 bp B0034 - 250 bp