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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

*Criobiology applied to plant crops

After a selection process we chose the Criobiology Project

The final design of our expresions moduls is as follow.

Next Notebook paper is our reference for primer's use, ligations and moduls Assembly.

The AFP (Antifreeze protein) was synthesized as follow

Week #1

06th September - 10th September 2010

Monday

We discussed and concluded the Igem’s vector (vial with green cover) is not enough.

First step transform only vector to get enough and begin ligations.

We are going to transform Psb1C3 from plate. For this:

• Prepare a stock of Cloramphenicol, Kanamicyn, Ampicilin solutions.

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

• Complete procedure for making quimio and electro-competent cells.

Tuesday

Completed competent cells and stored aliquots at -80 degrees each vial with 150μl.

We transformed DH5α cells with Psb1C3.

Wednesday

For a strange reason we have not transformats cells

today we are going to check out the Cloramphenicol

dose and try again the transformation with Psb1C3.

Thursday

In vector’s absence we have recived the primer’s synthesis

and proceed to amplify by PCR.

The amplification is correct and we have to looking for a nanodrop

to quantify DNA’s concentrations.