User:Daniel Ramirez/Notebook/UNAM Genomics Mexico 2011/2011/07/27

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χρόνος πέρασμα July 27th 2011


  • Double digestion for Standard Assembly of Bioparts E0430 and J04450 for the transcriptional terminator B0015 characterization construction.

  • Today I checked the digestion of the plasmids that carry the bioparts E0430 and J04450, these were digested only by EcoRI-HF. Lane #1 contain 1kb DNA ladder, lanes #2 and #3 contain E0430 digested by EcoRI-HF, lanes #4 and #5 contain J04450 digested by EcoRI-HF. The sizes of the plasmids that contain the bioparts are approximately 3 kb, the gel confirms the sizes. As it can be seen, there are another bands corresponding to undigested plasmids for lanes #3, #4 and #5.

Dramirez EcoRIdigestion E0430 J04450.jpg

  • I performed again the digestions but now with the other enzymes that are needed for Standard Assembly [1] using the same plasmid extraction from yesterday. To avoid partial digestion, the same amount of plasmidic DNA will be used to each digestion (corresponding to the DNA concentration of the E0430 #1 extraction). This is how the double digestions were carried out:

H20 12 μl
Buffer #4 3 μl
BSA 2 μl
EcoRI-HF 1.5 μl
XbaI(E0430)/SpeI(J04450) 1.5 μl
DNA 10 μl
Total 30 μl

  • The digestions were left overnight incubating at 37°C.

  • I went to talk to Maluye to ask her for the devise that induces microaerobiosis. She agreed to lend it to us. I also asked her the two questions that our collaborators from Estudios Latinoamericanos asked us; these were: What is the threeshold in nitrogen concentration in soil that causes the plant not to develop nodules and fix nitrogen. Maluye answered me that she didn't know, but she would help us seeking it in other sources. I will ask it to Esperanza Martínez. Is the plant somehow modified by the rhizobium in our project? Maluye told me that there isn't any evidence so far that proposes that the plant genome is modified during the simbiosis, that it is extremely difficult to modify the plant itself (hasn't been accomplished throughout these years).
  • I also continued helping Melissa Molho and Vladimir Muñoz in the Anderson collection of promoters plasmid extraction.