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 all documents ready to send them at the CONACyT.

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• Status: Work in progress
• Issues: None

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This biobricka are for the Diffusible signal oscillator and are intended to be assembled into the first Mexican biobrick.

First Mexican part!

• <bbpart>BBa_I0462</bbpart> available at well 11M at iGEM 2006 DNA1 1 in plasmid pSB1A2
• <bbpart>BBa_R0063</bbpart> available at well 9I at iGEM 2007 Parts Kit Plate 1 in plasmid pSB1A2

This biobrick is a Diffusible signal oscillator devise designed by the Pakrash group.

• <bbpart>BBa_I4204</bbpart> available at well 11O at iGEM 2007 Parts Kit Plate 1 in plasmid pSB1AK3

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 <bbpart>BBa_I0462</bbpart> and <bbpart>BBa_R0063</bbpart> have Amp resistance while <bbpart>BBa_I04204</bbpart> 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.

• The pasmid containing <bbpart>BBa_R0063</bbpart> should be around 2230bp
• The plasmid containing <bbpart>BBa_I0462</bbpart> should be around 3015bp

Then make the proper cuts in each plasmid with the proper endonucleases:

• The pasmid containing <bbpart>BBa_R0063</bbpart> should be cut with EcoRI and SpeI
• The plasmid containing <bbpart>BBa_I0462</bbpart> should be cut with EcoRI and XbaI

The cuts would then release <bbpart>BBa_I0462</bbpart> and it must be separated from pSB1A2 in order to ligate it with <bbpart>BBa_R0063</bbpart> there I don't know how to do that... perhaps another gel electrophoresis with very few DNA?