Team:UPO-Sevilla/Notebook/07 26


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July, 26th

Production Team

Paola Gallardo. Products of the first PCR reaction were analyzed by electrophoresis (0'8 %). We got four bands clearly-defined with the correct lengths (635 and 732 for gltD, 1206 and 312 for fecI-fecR). Purification from gel.

David Caballero. Plate analysis: colonies were presented in all plates except for fecA* and control plates. We verified the presence of fecI and PfecA parts in colonies by colony PCR reactions and 1.5% agarose gel electrophoresis analysis. In other way, to obtain fecA* part we digested by restriction enzymes the amplified by PCR part again, ligated it and transformed competent bacteria which remained overnight at 37ºC in LB+Km plate.

Assembly Team

We repeated PCR colony reaction because we hadn’t anything confirmed yet. We analyzed UPO1+UPO2 with Nhe1 but results were negative. Moreover, we did PCR colony reaction for the new UPO1+2 part. We are moving forward too slow. In addition we are waiting for the synthesized DNA... We hope it comes soon!

And again, digestion and ligation of UPO1 and UPO2 using the GINGO kit. Later, transformation in DH5α

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