Team:UTP-Panama/Experiments2

From 2011.igem.org

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(GaTech 2010 Biobrick BBa_k41000)
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== GaTech 2010 Biobrick BBa_k41000 ==
== GaTech 2010 Biobrick BBa_k41000 ==
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GaTech 2010 Biobrick BBa_K410000<br>
Objectives:<br>
Objectives:<br>
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1. Test the BioBrick operation to different substrates and temperatures:<br>
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1. Test the BioBrick operation to different substrates and temperatures:
a. Substrates: <br>
a. Substrates: <br>
i. LB <br>
i. LB <br>
ii. Minimal essential medium<br>
ii. Minimal essential medium<br>
iii. Saline solution<br>
iii. Saline solution<br>
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iv. Lunar land<br>
b. Temperatures:<br>
b. Temperatures:<br>
i. 8°C<br>
i. 8°C<br>
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ii. X°C<br>
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ii. 23°C<br>
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iii. 23°C<br>
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iii. 37°C<br>
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iv. 37°C<br>
2. Improve the characterization of the BioBrick tested in other substrates to study the performance of the promoter.<br>
2. Improve the characterization of the BioBrick tested in other substrates to study the performance of the promoter.<br>
3. Define the Biobrick promoter strength at the temperatures and selected substrates.<br>
3. Define the Biobrick promoter strength at the temperatures and selected substrates.<br>
Methodology<br>
Methodology<br>
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We chose 4 temperatures: <br>
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1. We used nine falcon tubes of 25 mL. Three tubes for every different substrate.<br>
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• 5-10°C<br>
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2. To each tube we added 10 ml of liquid LB, 10 mL of chloramphenicol (33μg/mL) and 500 µL of K2NO3 40 mM.<br>
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• 10-20°C<br>
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3. Tubes corresponding the different substrates were inoculated with BBa_K381001.<br>
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• 20-30°C<br>
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4. Tubes were incubated at 37, 8 and 23 ° C overnight.<br>
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• 37°C<br>
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These temperatures were selected so we could evaluate the response to cold shock in various ranges of our BioBrick. Since the lab equipments we had available couldn’t reach temperatures between 11-20°C this temperature was not used during the experience. Finally, the lab equipment allowed us to make the experiments in: 37°C, 23°C and 8°C.
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We also chose 3 different Growth Media: <br>
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• Saline Solution<br>
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• Liquid LB<br>
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• Minimal Media<br>
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These growth medias where selected to evaluate the response of our BioBricks in different metabolic pathways.
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We prepared 9 15mL Falcon Tubes, these tubes contained: <br>
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• First 3 (one for each temperature): 10mL of Saline Solution + a colony of the BioBrick.<br>
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• Second 3 (one for each temperature): 10mL of Minimal Media + a colony of the BioBrick.<br>
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• Lat 3 (one for each temperature): 10mL of Liquid Luria Broth + a colony of the BioBrick.<br>

Revision as of 03:41, 29 September 2011

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GaTech 2010 Biobrick BBa_k41000

GaTech 2010 Biobrick BBa_K410000
Objectives:
1. Test the BioBrick operation to different substrates and temperatures: a. Substrates:
i. LB
ii. Minimal essential medium
iii. Saline solution
iv. Lunar land
b. Temperatures:
i. 8°C
ii. X°C
iii. 23°C
iv. 37°C
2. Improve the characterization of the BioBrick tested in other substrates to study the performance of the promoter.
3. Define the Biobrick promoter strength at the temperatures and selected substrates.

Methodology
We chose 4 temperatures:
• 5-10°C
• 10-20°C
• 20-30°C
• 37°C
These temperatures were selected so we could evaluate the response to cold shock in various ranges of our BioBrick. Since the lab equipments we had available couldn’t reach temperatures between 11-20°C this temperature was not used during the experience. Finally, the lab equipment allowed us to make the experiments in: 37°C, 23°C and 8°C. We also chose 3 different Growth Media:
• Saline Solution
• Liquid LB
• Minimal Media
These growth medias where selected to evaluate the response of our BioBricks in different metabolic pathways. We prepared 9 15mL Falcon Tubes, these tubes contained:
• First 3 (one for each temperature): 10mL of Saline Solution + a colony of the BioBrick.
• Second 3 (one for each temperature): 10mL of Minimal Media + a colony of the BioBrick.

• Lat 3 (one for each temperature): 10mL of Liquid Luria Broth + a colony of the BioBrick.