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| Ter K2 ---> Terminator grown in medium with kanamycin. | | Ter K2 ---> Terminator grown in medium with kanamycin. |
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- | '''Measurements:''' | + | '''DNA concentration measurements:''' |
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- | DNA quantification with Qubit fluorometer (invitrogen). | + | DNA quantification with Qubit fluorometer (Invitrogen). |
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| Plasmid Terminator Concentrations: | | Plasmid Terminator Concentrations: |
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- | This was placed 3 hours at 37° C on the thermocycler. It takes 20 minutes at 80° C to deactivate enzymes. | + | This was placed for 3 hours at 37°C on the thermocycler. It takes 20 minutes at 80°C to deactivate enzymes. |
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- | PCR gDNA. Pseudomona (Rh1AB)
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- | II Test. Pair of primers:
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| + | Rh1AB(Pseudomona) PCR amplification: Second Test |
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| Primers: | | Primers: |
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| 2) 94° C 30 segs. | | 2) 94° C 30 segs. |
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- | 51° C 1 min.
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- | 72° C 3 min.
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| 3) 72° C 10 mins. | | 3) 72° C 10 mins. |
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| {{:Team:Panama/calendar2}} | | {{:Team:Panama/calendar2}} |
September 10
Ter A1 ---> Terminator grown in medium with ampicillin.
Ter A2 ---> Terminator grown in medium with ampicillin.
Ter K1 ---> Terminator grown in medium with kanamycin.
Ter K2 ---> Terminator grown in medium with kanamycin.
DNA concentration measurements:
DNA quantification with Qubit fluorometer (Invitrogen).
Plasmid Terminator Concentrations:
24.9ug/mL T1 (A1).
19.4ug/mL T2 (A2).
32.5ug/mL T3 (K1).
28.7ug/mL T4 (K2).
Agarose gel 1%.
Digestion Reactions of Plasmid,Reporter & Terminator.
Reporter
Plasmid
Terminator
This was placed for 3 hours at 37°C on the thermocycler. It takes 20 minutes at 80°C to deactivate enzymes.
Rh1AB(Pseudomona) PCR amplification: Second Test
Primers:
RhT-F1b5´ GTT TGC CTG TTC GAA AAT T 3´
RhT-2b 5´ CGA TAC GGC AAA ATC ATG G 3´
I. Resuspending the lyophilized primers for the solution stock (100 Plasmid u molar).
1). RhT-F1b
Tm 49.9° C, 34.1 nMoles= 6.10D260.
MW 5808.8
34.1 nMoles X 10= 341 uL H2O
Resuspending with 341uL H2O [ ]= 100 uMolar stock.
Work solution (10 uMolar)
2). RhT-2b
Tm= 51.7° C
MW= 5845.9
46.6 nMoles= 9.000260
46.6 nM x 10= 466uL H2O
Resuspending in 466uL H2O [ ]= 100 uMolar stock
Work solution (10 uM)
We tried with 3 differents volumes
1) 2.5uL [1 um] Final volume= 25uL
2) 1.25uL [0.5 um] Final volume= 25uL
3) 1uL [0.4 um] Final volume= 25uL
Thermocycler Program
1) 94° C 5 mins.
2) 94° C 30 segs.
51° C 1 min.
72° C 3 min.
3) 72° C 10 mins.
PCR Reaction 1
PCR Reaction 2
PCR Reaction 3
1% Gel Terminators (Extraction)
May
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| [http://2010.igem.org/wiki/index.php?title=Team:Panama/1_May_2010&action=edit 1]
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[http://2010.igem.org/wiki/index.php?title=Team:Panama/3_May_2010&action=edit 3]
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| [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]
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| [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/21_June_2010&action=edit 21]
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| [http://2010.igem.org/wiki/index.php?title=Team:Panama/25_June_2010&action=edit 25]
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[http://2010.igem.org/wiki/index.php?title=Team:Panama/28_June_2010&action=edit 28]
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July
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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]
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| [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]
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[http://2010.igem.org/wiki/index.php?title=Team:Panama/12_July_2010&action=edit 12]
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| [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]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/18_July_2010&action=edit 18]
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[http://2010.igem.org/wiki/index.php?title=Team:Panama/19_July_2010&action=edit 19]
| [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]
| [http://2010.igem.org/Team:Panama/24_July_2010 24]
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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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August
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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]
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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]
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[http://2010.igem.org/wiki/index.php?title=Team:Panama/16_August_2010&action=edit 16]
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[http://2010.igem.org/Team:Panama/23_August_2010 23]
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[http://2010.igem.org/wiki/index.php?title=Team:Panama/30_August_2010&action=edit 30]
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September
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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]
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| [http://2010.igem.org/wiki/index.php?title=Team:Panama/18_September_2010&action=edit 18]
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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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October
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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]
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[http://2010.igem.org/Team:Panama/18_October_2010 18]
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| [http://2010.igem.org/Team:Panama/23_October_2010 23]
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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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