August, 10th

Production Team

We repeated SDM reactions for fecA* (UPO28) and gltD** (UPO17) from the beginning. This time we used Taq expand enzyme instead of Pfu. In the results, we did not obtain any product of amplification in the first SDM’s PCR reaction. Next we made the second SDM’s PCR reaction using first former SDM’s PCR reaction products. Again the results were not satisfactory.

Assembly Team

Transformation of E. coli DH5α with ligations made the day before.

BamI-PstI digestions were analyzed by 1% agarose electrophoresis gel: colony 4 and 5 were positive.

The 2+28 and 28+3 day before transformation plates had not white colonies. Colonies from UPO7+2+28 plate were strange. We will repeat these devices using low copy vectors. We set up inocula of positive candidate colonies.

We made preparative digestion of UPO 4, 5 and 11 (in Mr. Gene vector) and ran them in 1% agarose gel. The spots were purified using GFX. Ligations with pSB1C3 and transformations were made.