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As summer proyect our experimental work began at 07 - july - 2009 using 2008 bioparts catalog, unfurtunly it dosen’t work properly, we had to much troubles to take out DNA and transform, E. Coli for this we had to delay and request 2009 catalog.

Our proyect uses 29 bioparts, 12 ligations and diferent strategys to make it functional as follow:

June

July

07-July-2009

We digest the follow biobriks with ECORI (E), SPEI (S), and XBAI (X).

Biobrick	Enzime restriction
R0079	E, S
F1610	E, X
B0034	E, S
C0078	E, X
C0079	E, X
B0015	E, X

We propose this restrictions for standar assembly for the digest we use a 17 hours incubation at 37º camera. Digest Mix:

ERCO RI - SPEI	Per reaction
Plasmidic DNA	3μl
Enzima ECORI	2μl
Enzima SPEI	2μl
Buffer NBE	2μl
BSA	0.5μl
H2O	10.5μl
Total	20μl

ECORI - XBAI	Per reaction
Plasmidic DNA	3μl
Enzima ECORI	2μl
Enzima XBAI	2μl
Buffer NBE	2μl
BSA	0.5μl
H20	10.5μl
Total	20μl

08-July-2009

We carried out the restrictions on a 10 wells agarosa gel 40 min. 3μl DNA 2μl Buffer. We have not DNA on the Gels maybe the DNA volume is so low. We try again with 20μl DNA 3μl Buffer We run on only Plasmidic DNA but we have not DNA.

09-July-2009

We start from cero, and take off DNA from the folder plateing R0079, C0178, C0179, C0060, B0034, I739001 biobriks, We left 16 hours in 37º camera.

10-July-2009

We don’t have transform cells in this point we have to delay and request 2009 catalog.

28-July-2009

With the 2009 catalog we retake the work and take on the biobriks we need. First of all we did a test to be sure that the biobriks worked properly for this we take a biobrik with RFP reporter (BBa_I3522) and plate, we incubate 17 hours in 37º camera.

29-July-2009

The transformation was successful we are sure the biobricks and method works and we proceed to take off our biobricks as next chart.

Number	Biobrick
2	BBa_R0079
3	BBa_F1610
4	BBa_K091146
5	BBa_K093005
6	BBa_K081016
7	BBa_EC840
8	BBa_K081009
9	BBa_R0051
10	BBa_J06800
11	BBa_Q04121
12	BBa_B0034
13	BBa_C0079
14	BBa_C0179
15	BBa_B0015
16	BBa_K081018
17	BBa_K116640

For transformation we use the same method as follow.

Elusion: 3μl DNA from the well, and get in Eppendorf 1.5 ml with competent cells. We use electoporation shock to transform and recovery in LB ampr 1 hour and a half then we plate on petri dishes and get incubate 17 hours.

31-July-2009

We plated Top 10 biobricks F1610, K091146, K081016, K081009, R0051, K081018 and get incube 17 hours

Team:IPN-UNAM-Mexico/Notebook