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Team:UC Davis/Notebook/Week 8
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--Monday 8/01/11--
Our transformations yielded colonies, but there were much more than expected on our vector control plates. We decided to PCR screen 4 colonies from each of the the ligations. After running them on a gel, it looks as though some of these colonies were successful ligations! At the end of the day, we cultured E0240+I13453 so that we could proceed with more construction tomorrow.
--Tuesday 8/02/11--
After miniprepping the cultured E0240+I13453, we set up a fast digest such that we could try a triple ligation. Our goal parts were: E0240+I13453+C0040, E0240+I13453+C0012, and E0240+I13453+C0051. We cut both E0240+I13453 and our repressor sequences as inserts and ligated them into a pSB1C3 backbone. We transformed and plated the ligation products.
--Wednesday 8/03/11--
Our triple ligation plates don't look very good. Most plates only have one colony while our vector control (pSB1C3) has about three. Today, our plan is to attempt to construct the same parts but by using our usual approach with fast digest enzymes. We digested, ligated, transformed, and plated these parts. Hopefully tomorrow we will see better results.
--Thursday--
--Friday--
