Team:UPO-Sevilla/Notebook/07 15


July, 15th

Production Team

Paola Gallardo. Using the ligation products we got the last day, we transformed competent bacteria and spread in LB+Cm plates once again. We hoped to get better results this time. New inocula of UPO16 (aspartate ammonia lyase) and UPO3 (rrnBT1-T7TE) were set up.

David Caballero. We continued without getting all parts, so we performed other PCR reactions for parts still not amplified. PCR reactions products were analyzed by 0.8% agarose gel electrophoresis and we realized presence of parts fecR*, PfecA, gltD** and fecI-fecR*. How it showed, the biggest parts, fecA* (2,3 kbp) and gltB (4,5 kbp), could not be obtained by PCR cycle we used.

Assembly Team

Today we’ve started with the UPO16+3 assemblies again.

After yesterday’s disappointment, we started over again digesting and ligating UPO 16+UPO 3. The GINGO kit arrived this morning, that’s why we hope to make ligation faster.

  1. Digestion and ligation of UPO3 and UPO16 using the GINGO kit. Later, transformation in DH5α
  2. E.Coli transformation with UPO2+13 and UPO1+19.
  3. Colony PCR of bacteria which have been growing all night

DryLab Team

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