Revision as of 01:32, 21 October 2009 by Cjdg (Talk | contribs)



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:

Month-icon.png June

Month-icon.png July



We made mini preped (protocol) and did the follow restrictions:

Number Biobrick Restriction
2 BBa_R0079 S & P
3 BBa_F1610 X & P
4 BBa_K091146 S & P
5 BBa_K093005 X &
6 BBa_K081016 X & P
7 BBa_EC840 X & P
8 BBa_K081009 X & P
9 BBa_R0051 S &P
10 BBa_J06800 X & P
11 BBa_Q04121 X & P
12 BBa_B0034 S & P
13 BBa_C0079 E & S
14 BBa_C0179 E & S
15 BBa_B0015 E & X
16 BBa_K081018 E & S
17 BBa_K116640 X & P
23 BBa_S03154 X & P
24 BBa_k145201 E & S


  • Prepared falcon tubes with 5ml agar liquid Ampr and inoculate.


  • Miniprep next clones: #16, #8, #23, #24, and prepare glycerol stocks.


  • Digest wit ECORI we use this method to check out if is the plasmid correct and begin ligations.


We start with ligations 15-13 (BBa_K266002) and 2-3 (BBa_K266000).

In this step we use 2 as backbone and 3 as insert, on the same 15 as backbone and 13 as insert, for 2-3 (BBa_K266000) we expect 3034 bp band as shown below, for 15-13 4074 bp


"Check plasmid and gel picutre below"

We carried out ligation 4-23 (BBa_K2660059) and consider unsuccessful, we proceed to repeat the transformation.


We did ECORI 3 hours restriction for 3, L 4-23 (BBa_K266005), 17, 3, 7 we see an inespecific band on 17 maybe a sobredigestion, we proceede to repeat this one.

"Check plasmid and gel picutre below"

12 -Agust-2009

We procede with 17_24 (Bba_K266001) ligation on this we going to use 17 as backbone and 24 as insert, we leave a 16 hours ligation in 14º camera.


We did transformation using heat shock then plate.


The transformation was successful and proceed to incubate tree colonys 17 hours in liquid LB medium.


We did midi prep for the ligation and carried out the plasmid is successful.

"Check plasmid and gel picutre below"


We going to proceed with J23100 – 11 (Bba_K266008) J23100 is a constitutive promoter and we going to use as backbone 11 as insert. We leave overnight ligation in 14º camera.

"Check plasmid and gel picutre below"


We did a transformation for thermic shock and plate, we expect colonys for midi


The transformation was unsuccessful we dont get colonys we going to try again.


We get Amplicon – L15-13 (BBa_K266003): Restriction digest 16 hours at 14º (overnight) then we will use thermic shock for cells transform.

L 2-3 (BBa_K266000) we’re doing ECORI 3 hours digest at 37º.

For BBa P1010 we take off the ccdB gene for this use PSTI and ECORI double restriction we going to use this part as chloramphenicol resistance vector .

For 24-17 BBa_K266001 we get this ligation and proceed to double digest

Month-icon.png September

Month-icon.png October

Banner footer UNAM2.jpg