Illinois/14 July 2009

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July 14

Hybrid Promoter

In lab we performed a miniprep of the inoculated colony from yesterday to isolate the DNA. It took some time to get access to the centrifuge as it was in use for much of the morning.

At the meeting with the professors they suggested that we take a look at the availabe GFP generator biobricks in the registry. It was also suggested that we would need DH5 alpha Z E. Coli cells if we are to use the lac promoter. It was also mentioned that about two plasmids transformed into a cell was standard and that we could probably get three.

Modeling

We miniprepped our transformed colonies to obtain DNA for BBa_K113009 and BBa_I0500. We also ran a gel on our PCR reactions on the pSB3K3 plasmid backbone. Both reactions yielded ~2kbp bands, when they were expected to be at 2.75kbp. We have decided to use this DNA anyway, since this is the second time bands have shown up at 2kbp.

We planned on digesting our DNA overnight, but our DNA concentrations were too low: between 5 and 30 ng/μL per sample. We have to find out what we should do to compensate for low concentrations.


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