Team:Panama/10 September 2010

From 2010.igem.org

iGEM Panama

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

alt text

Plasmid

alt text

Terminator

alt text

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

alt text

PCR Reaction 2

alt text

PCR Reaction 3

alt text

1% Gel Terminators (Extraction)

alt text

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]
[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]
[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]
[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]
[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]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/31_May_2010&action=edit 31]

June
M	T	W	T	F	S	S
	[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]
[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]
[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]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/21_June_2010&action=edit 21]	[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]	[http://2010.igem.org/wiki/index.php?title=Team:Panama/27_June_2010&action=edit 27]
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July
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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]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/12_July_2010&action=edit 12]	[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]	[http://2010.igem.org/wiki/index.php?title=Team:Panama/18_July_2010&action=edit 18]
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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]

August
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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]
[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]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/16_August_2010&action=edit 16]	[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]	[http://2010.igem.org/wiki/index.php?title=Team:Panama/22_August_2010&action=edit 22]
[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]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/30_August_2010&action=edit 30]	[http://2010.igem.org/wiki/index.php?title=Team:Panama/31_August_2010&action=edit 31]

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]
[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]
[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]
[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]
[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]

October
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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]
[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]
[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]
[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]