Team:UTP-Panama/Experiments3

Our RENBO Biobrick BBa_K672000

Objectives:
1. Test the BioBrick operation to different substrates and temperatures, using K2NO3 40 mM:
a. Substrates:
i. LB
ii. Minimal essential medium
iii. Saline solution
b. Temperatures:
i. 8°C
ii. 23°C
iii. 37°C
2. Characterize the expression of the promoters and determine the device operation.
3. Define the more efficient temperature and substrate for the operation of this device to survive at low temperatures and be able to measure nitrate.
Methodology

According to the characterization made by Team Edinburgh 2009, the Pyear promoter (BBa_K216005) used by Bristol 2010, works best in minimal media and complex media with a nitrate concentration of 40mM. Using this reference, we added 40mM of KNO3 to every growth media in the different temperatures for Bristol 2010 - BBa_K381001 and UTP-Panama 2011 – BBa_K672000.

1. We used nine falcon tubes of 25 mL. Three tubes for every different substrate.
2. To each tube we added 10 ml of liquid LB, 10 mL of chloramphenicol (33μg/mL) and 500 µL of K2NO3 40 mM.
3. Tubes corresponding the different substrates were inoculated with RENBO.