IGEM:UNAM-Genomics Mexico/2009/Notebook/H2 Pae/2011/08/03

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  • To repeat the conjugation of the conjugation of our plasmid pBRR1MCS-5, which was been done by Pualina, but she didn't use CaCl2 in the PY medium and this compund is neccesary for the proper grow of R. etli. This conjuation is done in order to analyze the plasmid stability on R. etli


1. I put to grow in 1 mL of liquid PY with CaCL2 a big pellet of: E. coli S17 carrying the pBBR1MCS-5(donor), the cultive was left 4 hours. It's also neccesary to have R. elti CE3 grown in this moment because the R. etli duplication is more slowly, Miguel gave me one cultive of grown R. etli.

2. In a steril test tube I put 100μL of the grown E. coli S17, 200μL R. etli CE3.

3. I took all the 300 μL from the last test tube and I put it in the center of a petri dish with solid PY(CaCl2), it was left over nigth at 30°C