October, 1st

Assembly Team

Different samples were run in a 0.8% or 1.5% agarose gel. UPO13+3, 12+2 and 1+2+16+3 were confirmed.

We have checked different preparations of 1+2 and 12+2. Different samples of 1+2 were digested using SspI-SpeI and these digestions were run in a 1.5% agarose gel. PCRs of 12+2 (several versions) and 12 were made and were run in a 0.8% polyacrilamide/TBE gel. UPO1+2 was confirmed; but UPO12+2 results were negative. UPO12+2 primers were designed.

Digestion of UPO 7, 7+3, 13+3 and 16+3 (2h, 37ºC) to clone them at pSB4K5. Digestions were run in an agarose gel and positive spots (all of them except 13+3) were isolated and purified using GFX. Ligation and transformation in DH5α.