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| | + | {{:Team:Panama/Header2}} |
| | ==='''August 9'''=== | | ==='''August 9'''=== |
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| - | High resolution agarose gel (for small fragments). For this we used two lanes (1 and 5) for the molecular weight. We used the Lamda DNA/Hind III marker from Biotools. We decided to run duplicates of the samples, so we have the Promoter and RBS on lanes 2 & 3; lane 4 is empty and once again, the Promoter and RBS are in lanes 6 & 7.
| + | '''More electrophresis''' |
| | | | |
| - | ''Ethydium Bromide staining:'' | + | High resolution 2% agarose gel (for small fragments). In this electrophoresis we used two (1 and 5) molecular weight markers, the Lamda DNA/Hind III marker from Biotools. We decided to load duplicates of the samples, so we have the Promoter and RBS on lanes 2 & 3; lane 4 is empty and once again, the Promoter and RBS are in lanes 6 & 7. |
| | + | |
| | + | After the electrophoresis was done, we proceed to stain the gel with ethydium bromide: |
| | + | ''Ethydium Bromide staining procedure:'' |
| | | | |
| | The '''stock concentration''' was at 10mg/ml. | | The '''stock concentration''' was at 10mg/ml. |
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| | The '''concentration for gel staining''' was 0.5ug/ml. | | The '''concentration for gel staining''' was 0.5ug/ml. |
| | | | |
| - | We used 15ul of Ethydium Bromide in 300ml of dH2O. We stained it covered for 30 minutes. Afterwards, we washed it in water for 10 minutes each time. | + | We used 15ul of Ethydium Bromide in 300ml of dH2O. We stained it covered for 30 minutes. Afterwards, we washed it in water for 15 minutes each time. |
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| - | [[Image:9-8.JPG|200px|thumb|left|alt text]] | + | [[Image:Cuadro9agosto2b.jpg|500px|thumb|left|alt text]] |
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| | + | [[Image:GEL9AGOSTOi.jpg|300px|thumb|right|alt text]] |
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| - | '''Digestion Repetitions:'''
| |
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| - | 1. Promoter (using iGEM restriction enzymes)
| |
| | | | |
| - | [[Image:Untitled2.jpg]]
| + | We weren't able to visualize the band of the small fragments of the digestion. We can only see the band that corresponds to the plasmid. It is possible that the intensity of the small bands is not enough for them to be seen. |
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| - | Buffer2 = 5ul
| |
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| - | BSA = 0.5ul
| |
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| - | EcoR1 = 1ul
| |
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| - | Spe1 = 1ul
| |
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| - | DNA = 17.98 ul
| |
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| - | dH2O = 24.52
| |
| | | | |
| - | '''Final volume = 50ul'''
| |
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| - | 2. Promoter (using INDICASAT restriction enzymes)
| |
| | | | |
| - | Buffer2 = 5ul
| |
| | | | |
| - | BSA = 0.5ul
| |
| | | | |
| - | EcoR1 = 1ul
| + | '''Restriction reaction Repetitions:''' |
| | | | |
| - | Spe1 = 1ul
| + | 1. Promoter (using NEB restriction enzymes) |
| | | | |
| - | DNA = 17.98 ul
| + | [[Image:Untitled2.jpg|150px|thumb|left|alt text]] |
| | | | |
| - | dH2O = 24.52ul
| |
| | | | |
| - | '''Final volume = 50ul'''
| + | 2. Promoter (using other restriction enzymes) |
| | + | |
| | + | [[Image:Untitled2.jpg|150px|thumb|left|alt text]] |
| | | | |
| | 3. RBS | | 3. RBS |
| | | | |
| - | Buffer2 = 5ul
| + | [[Image:Untitled2.1.jpg|150px|thumb|left|alt text]] |
| | + | |
| | + | |
| | + | |
| | + | The reason behind testing different batches of the same restriction enzymes for the promoter was to find out the reason why we didn't see anything when we ran electroforesis gels. We had thought that maybe the restriction enzymes were not doing their job, so we tried digesting the parts with enzymes that INDICASAT provided. |
| | | | |
| - | BSA = 0.5ul
| |
| | | | |
| - | Xba = 1ul
| |
| | | | |
| - | Pst1 = 1ul
| |
| | | | |
| - | DNA = 11.90ul
| |
| | | | |
| - | dH2O = 30.6ul
| |
| | | | |
| - | '''Final volume = 50ul'''
| |
| | | | |
| | | | |
| - | The reason behind testing different batches of the same restriction enzymes for the promoter was to find out the reason why we didn't see anything when we ran electroforesis gels. We had thought that maybe the restriction enzymes were not doing their job, so we tried digesting the parts with enzymes that INDICASAT provided.
| |
| | | | |
| | '''PsB1C3 Plasmid transformation (Stock):''' | | '''PsB1C3 Plasmid transformation (Stock):''' |
August 9
More electrophresis
High resolution 2% agarose gel (for small fragments). In this electrophoresis we used two (1 and 5) molecular weight markers, the Lamda DNA/Hind III marker from Biotools. We decided to load duplicates of the samples, so we have the Promoter and RBS on lanes 2 & 3; lane 4 is empty and once again, the Promoter and RBS are in lanes 6 & 7.
After the electrophoresis was done, we proceed to stain the gel with ethydium bromide:
Ethydium Bromide staining procedure:
The stock concentration was at 10mg/ml.
The concentration for gel staining was 0.5ug/ml.
We used 15ul of Ethydium Bromide in 300ml of dH2O. We stained it covered for 30 minutes. Afterwards, we washed it in water for 15 minutes each time.
We weren't able to visualize the band of the small fragments of the digestion. We can only see the band that corresponds to the plasmid. It is possible that the intensity of the small bands is not enough for them to be seen.
Restriction reaction Repetitions:
1. Promoter (using NEB restriction enzymes)
2. Promoter (using other restriction enzymes)
3. RBS
The reason behind testing different batches of the same restriction enzymes for the promoter was to find out the reason why we didn't see anything when we ran electroforesis gels. We had thought that maybe the restriction enzymes were not doing their job, so we tried digesting the parts with enzymes that INDICASAT provided.
PsB1C3 Plasmid transformation (Stock):
- LB plates with Cloranfenicol at a concentration of 33ug/ml.
- LB plates with Tetracycline at a concentration of 15ug/ml.
| May
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| [http://2010.igem.org/wiki/index.php?title=Team:Panama/1_May_2010&action=edit 1]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/2_May_2010&action=edit 2]
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| [http://2010.igem.org/wiki/index.php?title=Team:Panama/3_May_2010&action=edit 3]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/4_May_2010&action=edit 4]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/5_May_2010&action=edit 5]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/6_May_2010&action=edit 6]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/7_May_2010&action=edit 7]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/8_May_2010&action=edit 8]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/9_May_2010&action=edit 9]
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| [http://2010.igem.org/wiki/index.php?title=Team:Panama/10_May_2010&action=edit 10]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/11_May_2010&action=edit 11]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/12_May_2010&action=edit 12]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/13_May_2010&action=edit 13]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/14_May_2010&action=edit 14]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/15_May_2010&action=edit 15]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/16_May_2010&action=edit 16]
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| [http://2010.igem.org/wiki/index.php?title=Team:Panama/17_May_2010&action=edit 17]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/18_May_2010&action=edit 18]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/19_May_2010&action=edit 19]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/20_May_2010&action=edit 20]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/21_May_2010&action=edit 21]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/22_May_2010&action=edit 22]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/23_May_2010&action=edit 23]
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| [http://2010.igem.org/wiki/index.php?title=Team:Panama/24_May_2010&action=edit 24]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/25_May_2010&action=edit 25]
| [http://2010.igem.org/Team:Panama/26_May_2010 26]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/27_May_2010&action=edit 27]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/28_May_2010&action=edit 28]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/29_May_2010&action=edit 29]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/30_May_2010&action=edit 30]
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| [http://2010.igem.org/wiki/index.php?title=Team:Panama/31_May_2010&action=edit 31]
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| June
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| [http://2010.igem.org/wiki/index.php?title=Team:Panama/1_June_2010&action=edit 1]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/2_June_2010&action=edit 2]
| [http://2010.igem.org/Team:Panama/3_June_2010 3]
| [http://2010.igem.org/Team:Panama/4_June_2010 4]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/5_June_2010&action=edit 5]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/6_June_2010&action=edit 6]
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| [http://2010.igem.org/Team:Panama/7_June_2010 7]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/8_June_2010&action=edit 8]
| [http://2010.igem.org/Team:Panama/9_June_2010 9]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/10_June_2010&action=edit 10]
| [http://2010.igem.org/Team:Panama/11_June_2010 11]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/12_June_2010&action=edit 12]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/13_June_2010&action=edit 13]
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| [http://2010.igem.org/wiki/index.php?title=Team:Panama/14_June_2010&action=edit 14]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/15_June_2010&action=edit 15]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/16_June_2010&action=edit 16]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/17_June_2010&action=edit 17]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/18_June_2010&action=edit 18]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/19_June_2010&action=edit 19]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/20_June_2010&action=edit 20]
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| [http://2010.igem.org/wiki/index.php?title=Team:Panama/22_June_2010&action=edit 22]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/23_June_2010&action=edit 23]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/24_June_2010&action=edit 24]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/25_June_2010&action=edit 25]
| [http://2010.igem.org/Team:Panama/26_June_2010 26]
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| [http://2010.igem.org/wiki/index.php?title=Team:Panama/28_June_2010&action=edit 28]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/29_June_2010&action=edit 29]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/30_June_2010&action=edit 30]
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| July
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| M | T | W | T | F | S | S
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| [http://2010.igem.org/wiki/index.php?title=Team:Panama/1_July_2010&action=edit 1]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/2_July_2010&action=edit 2]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/3_July_2010&action=edit 3]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/4_July_2010&action=edit 4]
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| [http://2010.igem.org/wiki/index.php?title=Team:Panama/5_July_2010&action=edit 5]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/6_July_2010&action=edit 6]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/7_July_2010&action=edit 7]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/8_July_2010&action=edit 8]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/9_July_2010&action=edit 9]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/10_July_2010&action=edit 10]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/11_July_2010&action=edit 11]
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| [http://2010.igem.org/wiki/index.php?title=Team:Panama/13_July_2010&action=edit 13]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/14_July_2010&action=edit 14]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/15_July_2010&action=edit 15]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/16_July_2010&action=edit 16]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/17_July_2010&action=edit 17]
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| [http://2010.igem.org/Team:Panama/20_July_2010 20]
| [http://2010.igem.org/Team:Panama/21_July_2010 21]
| [http://2010.igem.org/Team:Panama/22_July_2010 22]
| [http://2010.igem.org/Team:Panama/23_July_2010 23]
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| [http://2010.igem.org/Team:Panama/26_July_2010 26]
| [http://2010.igem.org/Team:Panama/27_July_2010 27]
| [http://2010.igem.org/Team:Panama/28_July_2010 28]
| [http://2010.igem.org/Team:Panama/29_July_2010 29]
| [http://2010.igem.org/Team:Panama/30_July_2010 30]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/31_July_2010&action=edit 31]
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| [http://2010.igem.org/wiki/index.php?title=Team:Panama/1_August_2010&action=edit 1]
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| [http://2010.igem.org/Team:Panama/2_August_2010 2]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/3_August_2010&action=edit 3]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/4_August_2010&action=edit 4]
| [http://2010.igem.org/Team:Panama/5_August_2010 5]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/6_August_2010&action=edit 6]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/7_August_2010&action=edit 7]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/8_August_2010&action=edit 8]
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| [http://2010.igem.org/Team:Panama/9_August_2010 9]
| [http://2010.igem.org/Team:Panama/10_August_2010 10]
| [http://2010.igem.org/Team:Panama/11_August_2010 11]
| [http://2010.igem.org/Team:Panama/12_August_2010 12]
| [http://2010.igem.org/Team:Panama/13_August_2010 13]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/14_August_2010&action=edit 14]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/15_August_2010&action=edit 15]
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| [http://2010.igem.org/wiki/index.php?title=Team:Panama/17_August_2010&action=edit 17]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/18_August_2010&action=edit 18]
| [http://2010.igem.org/Team:Panama/19_August_2010 19]
| [http://2010.igem.org/Team:Panama/20_August_2010 20]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/21_August_2010&action=edit 21]
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| [http://2010.igem.org/Team:Panama/23_August_2010 23]
| [http://2010.igem.org/Team:Panama/24_August_2010 24]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/25_August_2010&action=edit 25]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/26_August_2010&action=edit 26]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/27_August_2010&action=edit 27]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/28_August_2010&action=edit 28]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/29_August_2010&action=edit 29]
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| [http://2010.igem.org/wiki/index.php?title=Team:Panama/31_August_2010&action=edit 31]
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| M | T | W | T | F | S | S
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| [http://2010.igem.org/wiki/index.php?title=Team:Panama/1_September_2010&action=edit 1]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/2_September_2010&action=edit 2]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/3_September_2010&action=edit 3]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/4_September_2010&action=edit 4]
| [http://2010.igem.org/Team:Panama/5_September_2010 5]
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| [http://2010.igem.org/wiki/index.php?title=Team:Panama/6_September_2010&action=edit 6]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/7_September_2010&action=edit 7]
| [http://2010.igem.org/Team:Panama/8_September_2010 8]
| [http://2010.igem.org/Team:Panama/9_September_2010 9]
| [http://2010.igem.org/Team:Panama/10_September_2010 10]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/11_September_2010&action=edit 11]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/12_September_2010&action=edit 12]
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| [http://2010.igem.org/wiki/index.php?title=Team:Panama/13_September_2010&action=edit 13]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/14_September_2010&action=edit 14]
| [http://2010.igem.org/Team:Panama/15_September_2010 15]
| [http://2010.igem.org/Team:Panama/16_September_2010 16]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/17_September_2010&action=edit 17]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/18_September_2010&action=edit 18]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/19_September_2010&action=edit 19]
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| [http://2010.igem.org/wiki/index.php?title=Team:Panama/20_September_2010&action=edit 20]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/21_September_2010&action=edit 21]
| [http://2010.igem.org/Team:Panama/22_September_2010 22]
| [http://2010.igem.org/Team:Panama/23_September_2010 23]
| [http://2010.igem.org/Team:Panama/24_September_2010 24]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/25_September_2010&action=edit 25]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/26_September_2010&action=edit 26]
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| [http://2010.igem.org/wiki/index.php?title=Team:Panama/27_September_2010&action=edit 27]
| [http://2010.igem.org/Team:Panama/28_September_2010 28]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/29_September_2010&action=edit 29]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/30_September_2010&action=edit 30]
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| [http://2010.igem.org/wiki/index.php?title=Team:Panama/1_October_2010&action=edit 1]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/2_October_2010&action=edit 2]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/3_October_2010&action=edit 3]
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| [http://2010.igem.org/Team:Panama/4_October_2010 4]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/5_October_2010&action=edit 5]
| [http://2010.igem.org/Team:Panama/6_October_2010 6]
| [http://2010.igem.org/Team:Panama/7_October_2010 7]
| [http://2010.igem.org/Team:Panama/8_October_2010 8]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/9_October_2010&action=edit 9]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/10_October_2010&action=edit 10]
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| [http://2010.igem.org/Team:Panama/11_October_2010 11]
| [http://2010.igem.org/Team:Panama/12_October_2010 12]
| [http://2010.igem.org/Team:Panama/13_October_2010 13]
| [http://2010.igem.org/Team:Panama/14_October_2010 14]
| [http://2010.igem.org/Team:Panama/15_October_2010 15]
| [http://2010.igem.org/Team:Panama/16_October_2010 16]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/17_October_2010&action=edit 17]
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| [http://2010.igem.org/Team:Panama/18_October_2010 18]
| [http://2010.igem.org/Team:Panama/19_October_2010 19]
| [http://2010.igem.org/Team:Panama/20_October_2010 20]
| [http://2010.igem.org/Team:Panama/21_October_2010 21]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/22_October_2010&action=edit 22]
| [http://2010.igem.org/Team:Panama/23_October_2010 23]
| [http://2010.igem.org/Team:Panama/24_October_2010 24]
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| [http://2010.igem.org/Team:Panama/25_October_2010 25]
| [http://2010.igem.org/Team:Panama/26_October_2010 26]
| [http://2010.igem.org/Team:Panama/27_October_2010 27]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/28_October_2010&action=edit 28]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/29_October_2010&action=edit 29]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/30_October_2010&action=edit 30]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/31_October_2010&action=edit 31]
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