Team:UPO-Sevilla/Notebook/09 07


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September, 7th

Production Team

David Caballero. We performed the first overlapping PCR fo SDM. We performed three reactions to be able of obtain fecI-fecR and fecR parts by mutagenesis. In each reaction the mix is:

37’5 ul H2O + 0,5ul UPO28* + 4 ul dNTP (10mM) + 5 ul Pfu buffer + 1 ul Pfu + 1 ul A-primer + 1 ul B-primer = 50 ul Vt

Next we performed a preparative electrophoresis with PCR’s products. 0,8% gel electrophoresis showed that we obtained amplification enough to mutagenize fecR, but not fecI-fecR composite part. We isolated from gel the parts amplified, products of the first PCR reaction needed to the next one.

Assembly Team

We analyzed our work and determined the most important biobricks to be built. Most of them are still uncompleted, but time is running out. A new distribution of the sources was done this week. I began with the digestion of the unavailable biobricks and the purification of available ones. We lost 2+6 (not digested). After that, we proceeded to join the biobricks: (1+2)+(7+2), 4+3, 4+ pSB1C3 by ligation (o/n)

Assay Team

We started with a chemotaxis assays series today. We studied some possible strategies we could follow in order to see chemotaxis in bacteria (quantitative and qualitative assays).

We designed the method to put into practice the capillary assay developed by Adler and we prepared the material to carry it out.

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