Team:Michigan/Safety

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Safety

1. Would the materials used in your project and/or your final product pose:

a. Risks to the safety and health of team members or others in the lab?
b. Risks to the safety and health of the general public if released by design or accident?
c. Risks to environmental quality if released by design or accident?
d. Risks to security through malicious misuse by individuals, groups or states?

a. Ethidum bromide is used as an intercalating agent and a dye when testing double stranded DNA. While used in highly dilute solutions, this compound possesses some acute toxicity and is handled in its stock form with nitrile gloves and under a fume hood. All liquid waste contacting this compound is disposed of in dedicated jugs, and solid waste collected in marked containers as required by the University of Michigan.

Hydrochloric acid (HCl) is used sparingly in 1M and 6M concentrations to clean DNA from cuvettes and purification columns. Concentrated solutions of HCl are generally handled with long sleeves, gloves and stored in either a fume hood with diaper pads or a locking, latching acid storage cabinet. All acid waste is collected in a separate, labeled disposal jug.

Sodium Hypochlorite is used in household concentrations as a means of destroying organisms and organic material before disposal. This compound is handled with gloves on a dedicated bench in small amounts.

Solutions and plates containing rich media such as LB, SOB, and SOC are frequently used for the growth of E.Coli for purification and DNA extraction. While the strains of E.Coli used are not pathogenic and listed as biosafety level 1 laboratory environment, leftover stocks and plates are still autoclaved and disposed of separately from other chemical waste.

Bunsen burners are used in conjunction with flammable 70% ethanol solutions for the purpose of sterilizing certain laboratory implements such as plate spreaders and inoculation pics; such methods are capable of causing burns if misused.

b. Recombinant strains of E.Coli containing resistance to various antibiotics are likely the greatest threat to the general public, as antibiotic resistance can potentially spread to wild-type bacteria through horizontal transfer. Antibiotics commonly used in the lab include ampicillin and kanamycin, both of which are used in public health to treat infections. The strains of E.Coli used in this process are non-pathogenic and pose little risk to those handling them, let alone public health

Hydrochloric acid in the concentrations used is potentially dangerous to the public if mishandled. HCl possesses both caustic and reactive properties and is capable of evolving gaseous chlorine on contact with certain compounds. Exposure of strong solutions of HCL to the general public would have unpredictable consequences as such solutions are so reactive.

c. Rich media such as LB, if released in the environment in large quantities, has the ability to disrupt ecosystems through the facilitation of abnormal bacterial and fungal growth.

Hydrochloric acid, if released into the environment, poses little long-term hazard due to its reactivity and subsequent short half-life. However, it still possess the potential to alter ecosystems by changing the chemical makeup of soils and groundwater if released in sufficient quantities.

Ethidium bromide is utilized in minuscule quantities and poses little environmental danger at its concentration.

d. Large amounts of concentrated hydrochloric acid can potentially be used to produce more toxic compounds such as gaseous chlorine and mustard gas and be used as weapons, if the individuals or states in question possess sufficient knowledge and equiptment. Bacteria in the lab are non-pathogenic and cannot be used to make bioweapons without extensive modification.

Please explain your responses (whether yes or no) to these questions.

Specifically, are any parts or devices in your project associated with(or known to cause):

- pathogenicity, infectivity, or toxicity?

- threats to environmental quality?

- security concerns?

2. If your response to any of the questions above is yes:

a. Explain how you addressed these issues in project design and while conducting laboratory work.

b. Describe and document safety, security, health and/or environmental issues as you submit your parts to the Registry.

3. Under what biosafety provisions will / do you operate?

a. Does your institution have its own biosafety rules and if so what are they? Provide a link to them online if possible.

b. Does your institution have an Institutional Biosafety Committee or equivalent group? If yes, have you discussed your project with them? Describe any concerns or changes that were made based on this review.

c. Will / did you receive any biosafety and/or lab training before beginning your project? If so, describe this training.

d. Does your country have national biosafety regulations or guidelines? If so, provide a link to them online if possible.

4. OPTIONAL QUESTION: Do you have other ideas on how to deal with safety or security issues that could be useful for future iGEM competitions? How could parts, devices and systems be made even safer through biosafety engineering?

1. Would any of your project ideas raise safety issues in terms of:

researcher safety,
public safety, or
environmental safety?

Overall, our project is very safe. If we wanted to allow the public to look at our cell patterns, we would have to take precautions to ensure that any viewers would be safe. We have discussed this with the company we get our microarrays from, and we are sure that the microarray slides themselves are safe to work with.

2. Do any of the new BioBrick parts (or devices) that you made this year raise any safety issues? If yes,

did you document these issues in the Registry?
how did you manage to handle the safety issue?
How could other teams learn from your experience?

There are no major safety issues involved with our part. Even though the zinc finger binding domain binds to double-stranded DNA, it will not mutate the DNA in any shape or form (unlike ethidium bromide that goes between the bases of DNA). Additionally our protein that contains the zinc finger is inert, no nuclease activity.

3. Is there a local biosafety group, committee, or review board at your institution?

If yes, what does your local biosafety group think about your project?
If no, which specific biosafety rules or guidelines do you have to consider in your country?

We take safety seriously on our team. All of our members are required to attend training sessions provided by the Occupational Safety and Environmental Health administration. These sessions deal with General Lab Safety and Autoclave Training. Our team has had meetings in past years with medical faculty to discuss biosafety, and we will plan another meeting this year.

4. Do you have any other ideas how to deal with safety issues that could be useful for future iGEM competitions? How could parts, devices and systems be made even safer through biosafety engineering?

All iGEM teams should go through mandatory lab safety training at a minimum. In order to make parts safer, we could require that each part pass the approval of a “safety committee.” This is not ideal however, because it would slow down the cloning process and would be expensive in terms of time and cost to implement. We could also prepare tests that each part could go through to ensure that they meet safety standards.

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-a. We do use materials in the lab that are potentially dangerous to us as team members handling them. One such example is ethidium bromide. Ethidium bromide is a potential mutagen because it works by inserting itself between the two strands of double-stranded DNA. While the amount of ethidium bromide kept in our lab is relatively small, it may still disrupt the replication of mitochondrial DNA.
+a. Ethidum bromide is used as an intercalating agent and a dye when testing double stranded DNA. While used in highly dilute solutions, this compound possesses some acute toxicity and is handled in its stock form with nitrile gloves and under a fume hood. All liquid waste contacting this compound is disposed of in dedicated jugs, and solid waste collected in marked containers as required by the University of Michigan.
-Concentrated hydrochloric acid (HCl) is another reagent we use in the lab that could be dangerous. HCl is an extremely strong acid that is dangerous either in mist or solution form. HCl is corrosive to human tissues and as such we handle is extremely carefully and as little as possible, diluting it before use if we can.
+Hydrochloric acid (HCl) is used sparingly in 1M and 6M concentrations to clean DNA from cuvettes and purification columns. Concentrated solutions of HCl are generally handled with long sleeves, gloves and stored in either a fume hood with diaper pads or a locking, latching acid storage cabinet. All acid waste is collected in a separate, labeled disposal jug.
-NaOCl, sodium hypochlorite, is another reagent we use that is potential dangerous. We use it in the form of household bleach to disinfect and clean our glassware, including glassware that had contained bacteria. Because we use a highly diluted form of bleach, it is no more dangerous than using it to clean in a household. We just take care to use gloves while washing glassware with the bleach and to not inhale heavily.
+Sodium Hypochlorite is used in household concentrations as a means of destroying organisms and organic material before disposal. This compound is handled with gloves on a dedicated bench in small amounts.
-b. While the materials listed in part a. could potentially be dangerous to the general public, we do not use any of them in high enough concentration to where accidentally releasing them could be dangerous to our surroundings. For example, ethidium bromide can be a mutagen for human DNA. However, ethidium bromide is sold in extremely small amounts and as such would not harm our surrounding community if released. The same is true for our hydrochloric acid and the sodium hypochlorite we use is the Chlorox brand that can be bought in stores for household consumption. If the two, hydrochloric acid and sodium hypochlorite, were combined, they could potentially create toxic chlorine gas. However, if this gas was released to the surrounding air outside our laboratory, the dangerous gas could diffuse throughout the air, reducing toxicity in one concentrated place.
+Solutions and plates containing rich media such as LB, SOB, and SOC are frequently used for the growth of E.Coli for purification and DNA extraction. While the strains of E.Coli used are not pathogenic and listed as biosafety level 1 laboratory environment, leftover stocks and plates are still autoclaved and disposed of separately from other chemical waste.
-c. There are some materials that we use that could potentially harm the environment if released. As stated in part c., chlorine gas could form if we allow our bleach and concentrated hydrochloric acid to mix. We keep these reagents separated to keep this from happening.
+Bunsen burners are used in conjunction with flammable 70% ethanol solutions for the purpose of sterilizing certain laboratory implements such as plate spreaders and inoculation pics; such methods are capable of causing burns if misused.
-d. Chlorine gas is dangerous and has been used as a weapon within the past 100 years. For example, it was used as a poison gas in World War I as well as by insurgent groups in Iraq in 2007. These events have occurred before our use of the two ingredients used to make chlorine gas, so it is clear that our use of them are not a cause of any new weapon.
+b.  Recombinant strains of E.Coli containing resistance to various antibiotics are likely the greatest threat to the general public, as antibiotic resistance can potentially spread to wild-type bacteria through horizontal transfer. Antibiotics commonly used in the lab include ampicillin and kanamycin, both of which are used in public health to treat infections. The strains of E.Coli used in this process are non-pathogenic and pose little risk to those handling them, let alone public health
+Hydrochloric acid in the concentrations used is potentially dangerous to the public if mishandled. HCl possesses both caustic and reactive properties and is capable of evolving gaseous chlorine on contact with certain compounds. Exposure of strong solutions of HCL to the general public would have unpredictable consequences as such solutions are so reactive.
+c.  Rich media such as LB, if released in the environment in large quantities, has the ability to disrupt ecosystems through the facilitation of abnormal bacterial and fungal growth.
+Hydrochloric acid, if released into the environment, poses little long-term hazard due to its reactivity and subsequent short half-life. However, it still possess the potential to alter ecosystems by changing the chemical makeup of soils and groundwater if released in sufficient quantities.
+Ethidium bromide is utilized in minuscule quantities and poses little environmental danger at its concentration.
+d. Large amounts of concentrated hydrochloric acid can potentially be used to produce more toxic compounds such as gaseous chlorine and mustard gas and be used as weapons, if the individuals or states in question possess sufficient knowledge and equiptment. Bacteria in the lab are non-pathogenic and cannot be used to make bioweapons without extensive modification.
 ''Please explain your responses (whether yes or no) to these questions.''