User:Daniel Ramirez/Notebook/UNAM Genomics Mexico 2011/2011/05/05

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χρόνος πέρασμα May 5th 2011

  • Some E. coli with the plasmid pBBRMCS5 resistant to gentamicin were left to grow overnight. They did grow on the petri dishes.

The negative controls didn't grow. Some bacteria were transposed on 5 ml of liquid medium with gentamicin and left to grow from morning to afternoon. I, along Melisa Rivas, proceded to extract the plasmid in order to have all three plasmids we wish to standarize. We followed the protocol for plasmid extraction. The plasmids were left isolated with TE-RNAse overnight.

  • We now posess the three plasmids, soon we will start with the standarization

In other news, I, along with Helena Reyes and Vladimir Muñoz, had a metting with David Romero. He explained us the methodology for the selection of the bacteria that will effectively have the constructions we wish them to have. With respect with the chromosome integration, he warned us about the region we chose in R. etli's chromosome to insert the synthesis; it seems to be tricky, it has an inverted segmental duplication that might prove to be difficult to deal with. The homology region we will add to the plasmids must be at least 500 bp because our constructions themselves have regions with high identity (meaning 100% identity) with etli's genome, fortunately, they are short in sequence (>250 bp, the NifH upstream region, among others).

  • Tomorrow we will proceed to clean our lab.



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