Team:UTP-Panama/Experiments4

From 2011.igem.org

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[[Team:UTP-Panama/Experiments| '''Experiment 1 ''']]  |
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[[Team:UTP-Panama/Experiments2| '''Experiment 2  ''']]  |
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[[Team:UTP-Panama/Experiments3| '''Experiments 3 ''']]  |
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[[Team:UTP-Panama/Experiments4| '''Experiments 4 ''']]  |
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[[Team:UTP-Panama/New Results| '''New Results''']]  |
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[[Team:UTP-Panama/Results|'''Results''']]|
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== New Experiments ==
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===Further Characterization===
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Objectives:
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1. Perform further experiments with our device
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2. Improve the characterization of THE RENBO
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3. Determine the relationship between the parameters we are studying (gene expression, temperature, substrate and thermal method)
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4. Obtain the proper functioning of our device
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Methodology:
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 We will use 5mL of substrates (liquid LB and MME)
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 We will have controls with and without antibiotics
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 We will use colonies of the following:
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o JM109 (just the strain with empty plasmids, control)
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o With RFP (control)
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o Bristol BBa_K381001
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o GaTech  BBa_K672000
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 Temperatures we used are:
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o 37ºC
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o Between 15ºC and 20ºC  (18ºC)
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 We will study growth using the two thermal methods:
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o Studying growth putting the colonies to grow directly at the chosen temperatures
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o Studying growth when the colonies have undergone a thermal shock @10ºC for 1 hour
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 For each temperature and BioBrick we wanted to get (in each experiment):
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o Growth curve
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o measurement of GFP expression
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o measurement of RFP expression (control)
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o measurement of heat production (thermometer)
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<div align="center">Experiments:</div>
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#1: All Colonies in LB and MM in blank, for references without antibiotics and without nitrate, @37ºC and @18ºC.
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#2: All Colonies in LB and MM with nitrate @37ºC and @18ºC.
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#3: All Colonies in LB and MM with antibiotics @37ºC and @18ºC.
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#4: All Colonies in LB and MM with antibiotics and nitrate @18°C and @37°C, first passing through thermal shock @10°C for 1 hour.
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#5: All Colonies in LB and MM with antibiotics and nitrate @18°C and @37°C, without thermal shock, direct growing.

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New Experiments

Further Characterization

Objectives: 1. Perform further experiments with our device 2. Improve the characterization of THE RENBO 3. Determine the relationship between the parameters we are studying (gene expression, temperature, substrate and thermal method) 4. Obtain the proper functioning of our device Methodology:  We will use 5mL of substrates (liquid LB and MME)  We will have controls with and without antibiotics  We will use colonies of the following: o JM109 (just the strain with empty plasmids, control) o With RFP (control) o Bristol BBa_K381001 o GaTech BBa_K672000  Temperatures we used are: o 37ºC o Between 15ºC and 20ºC (18ºC)  We will study growth using the two thermal methods: o Studying growth putting the colonies to grow directly at the chosen temperatures o Studying growth when the colonies have undergone a thermal shock @10ºC for 1 hour  For each temperature and BioBrick we wanted to get (in each experiment): o Growth curve o measurement of GFP expression o measurement of RFP expression (control) o measurement of heat production (thermometer)

Experiments:
  1. 1: All Colonies in LB and MM in blank, for references without antibiotics and without nitrate, @37ºC and @18ºC.
  2. 2: All Colonies in LB and MM with nitrate @37ºC and @18ºC.
  3. 3: All Colonies in LB and MM with antibiotics @37ºC and @18ºC.
  4. 4: All Colonies in LB and MM with antibiotics and nitrate @18°C and @37°C, first passing through thermal shock @10°C for 1 hour.
  5. 5: All Colonies in LB and MM with antibiotics and nitrate @18°C and @37°C, without thermal shock, direct growing.
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