Part:BBa_K847041:Experience

Assay
Cold
After transforming bacteria with our test plasmid, we grew approximately equal amounts of liquid culture of the negative control (see Test Plasmid) in LB+amp at 15°C, the lower limit of E. coli’s survival range. OD measurements were taken at time points t = 0 days and t = 9 days.
Desiccation
Liquid cultures of negative control, MntH, betAB, and otsAB transformed NEB5α E. coli were grown up over night at 37°C. After incubation, a dilution spot assay was conducted on each of the cultures to determine the density of live cells. Next, 15 5cm, round petri dishes were filled with 300uL of negative control bacteria, another 15 with transformants containing MntH, another 15 with transformants containing betAB, and another 15 with transformants containing otsAB. The petri dishes were allowed to desiccate while covered for 24 hours at 37°C while shaking. After the 24 hours period, each plate was resuspended with 1mL of fresh LB. A dilution spot assay was then conducted on each of the petri dish to determine the final density of live cells.

Results
Works for desiccation resistance. Refer to: http://2012.igem.org/Team:Stanford-Brown/HellCell/Desiccation Further testing needed for cold resistance. Refer to: http://2012.igem.org/Team:Stanford-Brown/HellCell/Cold

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