Notebook:Federico Castro M/2008/01/21


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Team reunion
Laboratorio de sistemática y biogeografía a las 11am.
 * We got to set ready all documents to send them at the CONACyT. I'm working on it.


 * Status: Work in progress
 * Two Biobricks successfully transformed, soon to be assembled ito the first mexican biobrick.
 * Issues: It's a rogue procedure.

Two biobricks were rehidrateated and sent to the IBT for further work.

This biobricka are for the Diffusible signal oscillator and are intended to be assembled into the first Mexican biobrick.
 * BBa_I0462 available at well 11M at iGEM 2006 DNA1 1 in plasmid pSB1A2
 * BBa_R0063 available at well 9I at iGEM 2007 Parts Kit Plate 1 in plasmid pSB1A2

The biobricks were rehidratated and saved for a trip to the IBT where Cuahutemoc is going to transform them and latter assemble the first Mexican biobrick.

It's very important to remember that BBa_I0462 and BBa_R0063 have Amp resistance while BBa_I04204 has resistance to Kan.

By mistake; Anglosaxon alphabet does not use ñ so we extracted a biobrick from the Biobrick at iGEM 2007 Parts Kit Plate 1 in the well 11P instead of the biobrick at well 11O. The biobrick was saved with all the stocks.

Cuahutemoc asked for the sequence of the plasmids in which biobricks are embeded so here is it

Further Work The following steep would be to assemble the two biobriks, I've never reached this steep before so I can only describe the process vaguely. For assembling the plasmids we first need to recover them from the bacteria, for that I guess one would need to grow a big population, extract the DNA and extract the plasmids with gel electrophoresis I guess thath we would be able to separate the plasmids from cromosomic DNA by the size of the plasmids. Then make the proper cuts in each plasmid with the proper endonucleases: The cuts would then release BBa_I0462 and it must be separated from pSB1A2 in order to ligate it with BBa_R0063 there I don't know how to do that... perhaps another gel electrophoresis with very few DNA?
 * The pasmid containing BBa_R0063 should be around 2230bp
 * The plasmid containing BBa_I0462 should be around 3015bp
 * The pasmid containing BBa_R0063 should be cut with EcoRI and SpeI
 * The plasmid containing BBa_I0462 should be cut with EcoRI and XbaI



(Study of Pakrash's oscillator)


 * Status: Work in progress
 * Issues: It's a rogue procedure.

One biobrick was extracted for further work at the IBT. The biobrick is a Difusible signal oscillator designed by the group of Pakrash and it will help us to understand the behavior of lactones and aiiA.


 * BBa_I4204 available at well 11O at iGEM 2007 Parts Kit Plate 1 in plasmid pSB1AK3