Illinois/13 July 2009

July 13
Hybrid Promoter

Today in lab we inoculated a colony of the cells that we transformed the biobricks into yesterday.

Concerns: We want the sRNA gene on a high copy plasmid and the target sequence on low copy plasmid. This could require up to 7 different plasmids to transform into the cell. We need to look into the feasibility of this.

Other things to consider:

- Can we put all of the sRNA expressing genes and their promoters on one plasmid(with all the target sequences and the genes that go with)? would this be too many ligations? How would we tell if they all worked? Run a gel?

- Would we co-transform the plasmids or do them separately?

Modeling

The transformation of BBa_K113009 yielded colonies, whereas the transformation of BBa_I0500 did not. We will have to redo the transformation of BBa_I0500.

We ran a PCR to amplify the pSB3K3 plasmid backbone for testing our promoters. For some reason, the gel we ran indicated a 6kbp band when it should have been 2.75kbp.

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