Experiments | Paris_Bettencourt

Impact on Growth of electroshock

Abstract

This experiment aim to investigate the parameters of electroshocking for our Electro-Genetic promoter screening experiment where we will electroshock bacteria's from the [E. coli Promoter collection](https://horizondiscovery.com/en/non-mammalian-research-tools/products/e-coli-promoter-collection)[1].

Indeed, exposure to electrical current can be lethal to organisms, we wanted to elucidate if exposure to electrical current would have an effect on growth.

We used a custom made [High throughput Electro Actuator (HTEA)](https://www.notion.so/f0034928f1414de6a9a001c32dd2694f) to test the impact on growth of exposure to AC Current

Our result show that none of the current within the range explored killed the bacterial population as growth is recovered after a small recovery phase.

We witness that the final OD vary depending on the shock

Materials and methods

As this experiment aim is to evaluate electroshock condition, stayed consistant with the strained used in [1] and worked with E. coli MG1655. We grew the bacteria's overnights on M9 (0.4% Glucose, complete Amino Acids).

In the morning, we diluted the cells into fresh M9 Media to a ratio of 1:100.

From a previous experiment[2], we determined that mid-log phase (OD600 = 0.4) was attained after 4 hours of growth . We waited 4 hours to perfom the electroshocking experiment

We used the HTEA in conjunction with the AC Dispatcher (ACD) and the Electro Planner to shock the bacteria with AC current of different amplitudes (from 0.2volts to 11 volts) .

We used the MHS2300A dual function generator and the MHS2300A Python library to communicate

Experiments were done in triplicates

High throughput Electro Actuator (HTEA)

Results

We observe that all population recovered growth after a slight lag that vary from conditions to conditions (Figure 1)

We also witness that the final OD reached vary depending on the experimental conditions. Interrestingly, the relationshionship between ac current applied and final OD is non linear (Figure 2)

Future Work

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References

1. Zaslaver, A. et al. (2006) ‘A comprehensive library of fluorescent transcriptional reporters for Escherichia coli’, Nature Methods, 3(8), pp. 623–628. Available at: https://doi.org/10.1038/nmeth895.