Revision as of 06:58, 21 October 2009 by Eniak (Talk | contribs)




  • Digestion of the following biobricks were done with EcoRI (E), SpeI (S), and XbaI (X).
Biobrick Enzyme restriction
R0079 E, S
F1610 E, X
B0034 E, S
C0078 E, X
C0079 E, X
B0015 E, X

  • We propose this restrictions for standard assembly for the digestion we did a 17 hours incubation at 37°C.
  • Digest Mix:
EcoRI - SpeI Per reaction
Plasmidic DNA 3μl
Enzyme EcoRI 2μl
Enzyme SpeI 2μl
Buffer NBE 2μl
BSA 0.5μl
H2O 10.5μl
Total 20μl
EcoRI - XbaI Per reaction
Plasmidic DNA 3μl
Enzyme EcoRI 2μl
Enzyme XbaI 2μl
Buffer NBE 2μl
BSA 0.5μl
H20 10.5μl
Total 20μl


  • We made the restrictions and put them on an agarose gel with 10 wells for 40 min.
  • We used 3μl DNA plus 2μl Buffer.
  • We didn't get any DNA from the gel so maybe the DNA volume is too low. We tried again with 20μl DNA plus 3μl Buffer
  • Run on only Plasmidic DNA but we have no DNA.


  • Starting again, and take off DNA from the folder plating R0079, C0178, C0179, C0060, B0034, I739001 biobriks,
  • Left 16 hours in 37ºC.


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


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


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.
  • Put it on a Eppendorf Vial 1.5 ml with competent cells.
  • Electoporate shock to transform and recovery in LB ampr 1 hour and a half.
  • Plate on petri dishes and incubate 17 hours.


  • Plated Top 10 biobricks F1610, K091146, K081016, K081009, R0051, K081018 and incubate during 17 hours

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