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Team:IPN-UNAM-Mexico/Notebook
From 2009.igem.org
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<h1>[[Image:Month-icon.png | 50px]] July </h1> | <h1>[[Image:Month-icon.png | 50px]] July </h1> | ||
| - | '''''07-July-2009''''' | + | ==='''''07-July-2009'''''=== |
We digest the follow biobriks with ECORI (E), SPEI (S), and XBAI (X). | We digest the follow biobriks with ECORI (E), SPEI (S), and XBAI (X). | ||
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| + | ---- | ||
| + | ---- | ||
| - | '''''08-July-2009''''' | + | ==='''''08-July-2009'''''=== |
We carried out the restrictions on a 10 wells agarosa gel 40 min. | We carried out the restrictions on a 10 wells agarosa gel 40 min. | ||
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We run on only Plasmidic DNA but we have not DNA. | We run on only Plasmidic DNA but we have not DNA. | ||
| - | '''''09-July-2009''''' | + | ---- |
| + | ---- | ||
| + | |||
| + | ==='''''09-July-2009'''''=== | ||
We start from cero, and take off DNA from the folder plateing R0079, C0178, C0179, C0060, B0034, I739001 biobriks, | 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. | 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''''' | + | ==='''''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. | 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''''' | + | ==='''''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. | The transformation was successful we are sure the biobricks and method works and we proceed to take off our biobricks as next chart. | ||
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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. | 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''''' | + | ==='''''31-July-2009'''''=== |
We plated Top 10 biobricks F1610, K091146, K081016, K081009, R0051, K081018 and get incube 17 hours | We plated Top 10 biobricks F1610, K091146, K081016, K081009, R0051, K081018 and get incube 17 hours | ||
Revision as of 17:28, 20 October 2009
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
August
September
October
