Team:UPO-Sevilla/Notebook/07 20


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July, 20th

Production Team

Paola Gallardo. Purification of plasmids from inocula with gltD** and fecI-fecR* (we accidentally destroyed fecI-fecR*-PfecA and fecR*-PfecA inoculPG), digestion of these plasmids with restriction enzymes EcoRI and PstI. We ran and electrophoresis with the digestion products and we checked if we have isolated colonies that contain these plasmids. Setting up of new inocula of E. coli bacteria with fecR*, PfecA, fecI-fecR*-PfecA y fecR*-PfecA; and finally, and isolation of the same bacterias in LB+Cm plates. We set up inocula of bacterias with fecR, PfecA, fecI-fecR-PfecA and fecR-PfecA.

David Caballero. Analysis of previous PCR reactions in 0.8% agarose gel electrophoresis. We had positives results for each amplified part: fecA* and gltB. The new parts were purified, digested and ligated in vectors with Cm resistance. Next we transformed competent bacteria with these parts and other ones: fecI, PfecA, fecI-fecR*, fecA* and gltB. We cultivated them in LB+Cm plates overnight to 37º.

Assembly Team

Digestion of UPO16+3 to check if they had the plasmid. Now we have another new biobrick (UPO16+3)!

Transformation plates of UPO1+19 and UPO2+13 were quite weird... Religation plate had about 100 colonies (we expected not to have anything). Ligation of UPO 2+3 had 1 colony and UPO1+19 had anything. We did colony PCR again to check that we marked well every plate. The gel said that we weren’t wrong and that maybe we have a colony with the right plasmid of UPO2+13. We put inocula of UPO2+13 and spread in a plate. Maybe ligation are giving problems because of the heat of the lab.

DryLab Team

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