IGEM:Cambridge/2008/Notebook/Turing Pattern Formation/2008/08/14

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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 plasmids.

Expected VF2-VR fragment sizes:

I13522 - ~2.4 kb 

E0040 -  958 bp
R0040 -  292 bp
B0034 -  250 bp



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