Team:Mexico-UNAM-CINVESTAV/Notebook/Week One

From 2010.igem.org

(Difference between revisions)
(After a selection process we chose the Criobiology Project)
Line 34: Line 34:
==''Monday''==
==''Monday''==
-
==='''We discussed and concluded the Igem’s vector (vial with green cover) is not enough.'''===
+
==='''We discussed and concluded that the Igem’s vector quantity  is not enough.'''===
-
==='''First step transform only vector to get enough and begin ligations.'''===
+
==='''First step:  Transform using only the vector to get enough material. '''===
-
==='''We are going to  transform Psb1C3  from plate.  For this:'''===
+
==='''We are going to  transform Psb1C3  from plate.  To do  this:'''===
-
*'''Prepare a stock of Cloramphenicol, Kanamicyn, Ampicilin solutions.'''
+
*''' We prepared  stock solutions of Cloramphenicol, Kanamicyn, Ampicilin.'''
*''' 5 LB agar and 35ug/ml cloramphenicol plates made up . '''
*''' 5 LB agar and 35ug/ml cloramphenicol plates made up . '''
-
*'''Complete procedure for making quimio and electro-competent cells.'''
+
*''' We completed the procedure to make quimio and electro-competent cells.'''
==''Tuesday''==
==''Tuesday''==
-
==='''Completed competent cells and stored aliquots at -80 degrees each vial with 150μl.'''===
+
==='''Finished competent cells and stored in aliquots in containers vials at -80 degrees.'''===
==='''We transformed DH5α cells with Psb1C3.'''===
==='''We transformed DH5α cells with Psb1C3.'''===
Line 57: Line 57:
==''Wednesday''==
==''Wednesday''==
-
==='''For a strange reason we have not transformats cells'''===  
+
==='''We have not obtained  transformats cells.'''===  
-
==='''today we are going to check out the Cloramphenicol'''===  
+
==='''We checked out the Cloramphenicol'''===  
-
==='''dose and try again the transformation with Psb1C3.'''===
+
==='''dose and attempt again the transformation using Psb1C3.'''===
==''Thursday''==
==''Thursday''==
-
==='''In vector’s absence we have recived the primer’s synthesis'''===  
+
==='''The primer’s arrived ''===  
-
==='''and proceed to amplify by PCR.'''===
+
==='''and we proceed to amplify the modules by PCR.'''===
[[Image:PCR.gif]]
[[Image:PCR.gif]]
-
==='''The amplification is correct and we have to looking for a nanodrop'''===  
+
==='''The amplification was ok and to quantify DNA concentrations'''===  
-
==='''to quantify DNA’s concentrations.'''===
+
==='''we had looking for a nano spectrophotometer.'''===

Revision as of 20:40, 26 October 2010



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:

caption


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


Primers1.JPG

The AFP (Antifreeze Protein) was synthesized as follow:

Vector001.jpg

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.

PCR.gif

The amplification was ok and to quantify DNA concentrations

we had looking for a nano spectrophotometer.