Safe Project Design

Does your project include any safety features? Have you made certain decisions about the design to reduce risks? Write about them here! For example:

• Choosing a non-pathogenic chassis
• RAINmakers are employing 5-alpha competent E. coli cells (DH5-Alpha strain). According to the Globally Harmonized Systems (GHS), this organism is not a dangerous substance or mixture. Per Centers for Disease Control and Prevention (CDC), this organism can be handled at Biology Safety Level One (BSL-1). The CDC goes on to state that 5-alpha competent E. coli cells present no risk of disease in immunocompetent adult humans and no hazard to laboratory personnel and the environment. There are certain first aid measures that must be taken if the 5-alpha E. coli cells come in contact with lab personnel. If the cells come in contact with skin, washing with soap and water is sufficient. If there is eye contact, rising thoroughly with water for 15 minutes is imperative, as well as consultation of a physician. In the case of inhalation, exposure to fresh air is all that is necessary. All members of Tacoma RAINmakers are versed in these simple first aid measures. In the case of accidental release into the environment, cells should be soaked up with inert absorbent material, transferred to a properly labelled container, and disposed of as hazardous waste. Still, it is important to recall that this organism presents no environmental threat.
• In lab, the first step for Tacoma RAINmakers is cloning our construct. We digested our vector, ArsR gene, and chromoproteins. Since ArsR and our chromoprotein make up the insert portion of the construct, they must be ligated. Then, the newly created insert is ligated into our vector, forming our full construct. The construct DNA is added to 5-alpha competent E. coli cells and grown in LB broth. At this point, the RAINmakers would "mini-prep" our E. coli cells using cell-free lysate kits. Now that the desired DNA material has been produced, we are able to proceed to our experiments. One such experiment will involve pipetting our circuit DNA into our paper ticket, exposing the ticket to arsenic solution, incubating, and qualitatively assessing chromoprotein production. Specific aspects of this experiments that must be optimized include incubation time, DNA quantity, and reagent proportions. Additionally, RAINmakers will empirically determine the maximum and minimum amount of arsenic concentration our biosensor is able to detect. Once our sensing mechanism is functional with known quantities of arsenic, we will test our ticket with environmental samples (i.e. water, diluted soil). All parts mentioned are already in the registry.