August, 10th

Assembly 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.

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

We analyzed BamI-PstI digestions in 1% agarose gel: colony 4 and 5 are positive.

The 2+28 and 28+3 day before transformation plates have not white colonies. 7+2+28 colonies are strange. We will repeat these devices using low copy vectors. We set up inocula of positive candidate colonies.

Preparative digestion of UPO 4, 5 and 11 (in Mr. Gene vector) and 1% agarose gel. GFX purification. Ligation with pSB1C3 and transformation.