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From 2010.igem.org

As summer project our experimental work began in August, after intensive brainstorming we finnaly decide

between two options.

*Inmunoresponse using Synthetic Biology

*Cryobiology applied to plant crops

After a selection process we chose the Cryobiology Project

The final design of the expressions modules were as follow:

Next Notebook sheet is our reference for primers and ligations and for modules Assembly:

The AFP (Antifreeze Protein) was synthesized as follow:

Week #1

06th September - 10th September 2010

Monday

We discussed and concluded that the Igem’s vector quantity is not enough.

First step: Transform using only the vector to get enough material.

We are going to transform Psb1C3 from plate. To do this:

• We prepared stock solutions of Cloramphenicol, Kanamicyn, Ampicilin.

• 5 LB agar and 35ug/ml cloramphenicol plates made up .

• We completed the procedure to make quimio and electro-competent cells.

Tuesday

Finished competent cells and stored in aliquots in containers vials at -80 degrees.

We transformed DH5α cells with Psb1C3.

Wednesday

We have not obtained transformats cells.

We checked out the Cloramphenicol

dose and attempt again the transformation using Psb1C3.

Thursday

'The primer’s arrived

and we proceed to amplify the modules by PCR.

The amplification was ok and to quantify DNA concentrations

we had looking for a nano spectrophotometer.