Team:UPO-Sevilla/Notebook/07 15
From 2010.igem.org
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.
- Digestion and ligation of UPO3 and UPO16 using the GINGO kit. Later, transformation in DH5α
- E.Coli transformation with UPO2+13 and UPO1+19.
- Colony PCR of bacteria which have been growing all night
DryLab Team
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