Team:TCU Taiwan/Safety

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Safety

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ENJOY THE iGEM!

All team member has to take lab safety training.

No eating in the lab.

Safety Form

1.Safety Training

a)Have your team members received any safety training yet?

Yes, we have already received safety training.

b)Please briefly describe the topics that you learned about in your safety training.

1.Universal lab safety rules: wearing lab coat and glasses, washing hands, keeping work area uncluttered, etc.
2.Know the location of fire extinguisher, eyewash station, chemical spill kits and else
3.First aid lesson
4.Help yourself: memorizing emergency safety telephone number

2.Local Rules and Regulations

a)Who is responsible for biological safety at your institution?

Environmental Health and Safety Center in Tzu Chi University.

3.About Our Lab

a)What is the Safety Level of your lab? (Use the WHO numbering system, in which Level 4 is used for the most dangerous organisms.)

Level 2 (moderate risk, ≌ WHO BSL 2)

b)According to the WHO Biosafety Manual, what is the BioSafety Level rating of your lab?

Laminar flow hood/biosafety cabinet with open front

Cute BoBo looks back with a smile.

c)What personal protective equipment do you use in your lab? Please check all that apply.

Lab coats

Gloves

Safety glasses / goggles

Surgical masks

d)How do you dispose of biological waste?

Liquid waste will be collected first and then sent to our school's liquid waste storing house.
Other waste will be collected into Biohazard waste disposal.

4.The Organisms and Parts

Species name (including strain) Risk Group Risk Group Source Disease risk to humans? Part number/name Natural function of part How did you acquire it? How will you use it?

TCU Taiwan Safety2014 Spreadsheet


Species name	Risk Group	Risk Group Source	Disease risk to humans?	How did you acquire it?	How will you use it?
E.coli top 10	1	WHO	do not cause disease in healthy adult humans.	Acquired from our lab	Competent cell for our plasmid
E.coli JM101	1	WHO	do not cause disease in healthy adult humans.	Acquired from another lab in our school	Model organism for infection by M13KO7
E.coli DH5α	1	WHO	do not cause disease in healthy adult humans.	Acquired from our lab	Competent cell for our plasmid
E.coli NEB10 β	1	WHO	do not cause disease in healthy adult humans.	Acquired from igem HQ	Competent cell for our biobrick
M13KO7 hlper phage	1	WHO	do not cause disease in healthy adult humans.	Bought from NEB	Vector for our system

Remember to change the filename of your spreadsheet! Put your team's name in place of "TeamName".

Species name (including strain): For an organism, give the scientific name of the species. Include a strain name or number (such as "K-12" for E. coli K-12) if there is one. For a part, give the name and strain of the organism that the part originally came from.
Risk Group: Give the Risk Group of the organism in column A. You may use a categorization according to your home country, according to the USA, or according to the WHO. If the organism falls into an 'in-between' or special category such as 2+ or 2-Agricultural, explain this category in the Notes column. If you cannot find any Risk Group categorization for this organism, write "N/A" and explain in the Notes column. (Multicellular organisms generally do not have a Risk Group.)
Risk Group Source: Cite the source from which you obtained the Risk Group information. See Risk Group Guide for recommended sources. If you got the information from the Canadian PSDS, from the NIH Guidelines, or from the DSMZ catalogue, you may simply write "PSDS", "NIH", or "DSMZ". Otherwise, please give a web link or a full citation for your source.
Disease risk to humans?: Does this organism cause any disease in humans? If yes, what disease does it cause?
Part number/name: For a part: If it has a Registry part number (like BBa_XXXXX), write that number. If it has no Registry part number, give a short name for the part. (For example: "Actin", "Alcohol Dehydrogenase".) For a whole organism, leave this column blank.
Natural function of part: For a part: Briefly describe what the part does in its parent organism. (If it is an enzyme, what reaction does it catalyze? If it is a receptor, what molecules does it bind to? Etc.) For a whole organism, leave this column blank.
How did you acquire it?: Describe how you acquired the organism/part. If you have not acquired it yet, describe how you plan to acquire it. (For example: did you receive the part DNA from another lab? Did you order the part DNA from a synthesis company? Did you use PCR to isolate the part from genomic DNA of its parent organism? Did you order the cell line from a company?)
How will you use it?: Describe how you are using the organism/part in the lab. (For example: "This organism is our chassis." "This part senses when the cells are exposed to glucose." "This organism is the source for a part that we are isolating by PCR." "This part produces the toxin which our bio-sensor is designed to detect.")
Notes: Use this column to give any additional information that is necessary.

5.Risks of Our Project Now

Please describe risks of working with the biological materials that you are using in your project. If you are taking any safety precautions (even basic ones, like rubber gloves), that is because your work has some risks, however small. Therefore, please discuss possible risks and what you have done to minimize them, instead of simply saying that there are no risks at all.

a)Risks to the safety and health of team members, or other people working in the lab:

Some reagents we use are poison for human. Such as EtBr, chloroform, etc.

b)Risks to the safety and health of the general public (if any biological materials escaped from your lab):

As our mechanism includes phage therapy, there might be some dangerous for injecting phage into human body.

c)Risks to the environment (from waste disposal, or from materials escaping from your lab):

CRISPR system may destroy bacteria's genome, thus break the balance of bacteria's population.

We try to engineer our gRNA to make it target only antibiotics resistant gene.

d)Risks to security through malicious mis-use by individuals, groups, or countries:

May used to cause unnecessary bacteria death in environment, result in break of bacteria balance.

e)What measures are you taking to reduce these risks?

1.We use helper phage and phagemid for phage display.
2.We try to engineer our gRNA to make it target only antibiotics resistant gene.

6.Risks of Our Project in the Future

What would happen if all your dreams came true, and your project grew from a small lab study into a commercial/industrial/medical product that was used by many people? We invite you to speculate broadly and discuss possibilities, rather than providing definite answers. Even if the product is "safe", please discuss possible risks and how they could be addressed, rather than simply saying that there are no risks at all.

a)What new risks might arise from your project's growth?

Also, what risks might arise if the knowledge you generate or the methods you develop became widely available?

Injecting phage into human body may cause some unknown danger.
May speed up E.coli to develop "resistance" to phage.

b)Does your project currently include any design features to reduce risks? Or, if you did all the future work to make your project grow into a popular product, would you plan to design any new features to minimize risks?Such features are not required for an iGEM project, but many teams choose to explore them.

We thought to change phage's coat protein so we can access our phage's infection ability. But this is beyond our current project.

7.Health Worker

Emergency Medical Technician (EMT) are terms used in some countries to denote a health care provider of emergency medical services.EMTs are clinicians, trained to respond quickly to emergency situations regarding medical issues, traumatic injuries and accident scenes.

House,one of our team members,is a Emergency Medical Technician.He got the EMT-1(EMT-B) Licence and he volunteered for Hualien County Fire Department.He can handle every medical emergencies for team members in daily life.

8.Project safety

As an environment project, the mean idea of our project’s safety is “not kill them but remove their resistance”, so here are some points.

We choose phage as our vector for the CRISPR system because of the high specificity. Each kind of phage will only infect one kind of bacteria efficiently.

To avoid the co-effect of phage infection, we choose the “phagemid-helper phage” system for phage display. This system creates progeny who cannot produce next generation along. Thus, our project will not cause additional phage remaining in the environment.

In our experiment, we choose M13 phage as model. This filamentous phage will make its genome a double strand plasmid in host bacteria’s cytosol. Therefore, M13 phage is not either a lytic or lysogenic phage, its release will not cause host cell’s death.

We design our CRISPR’s target as antibiotics resistance gene. So it will not influence bacteria’s growth as long as we don’t add antibiotics. We did this because we want the situation is always under control. (We DO NOT want to simply kill those bacteria, we just want to remove their resistance and make them under our control.)

TCU_Taiwan iGEM Laboratory Practices


1.	Don’t eat, drink, prepare or store food, smoke, handle contact lenses or apply cosmetics in any laboratory.
2.	Know where the nearest eyewash, safety shower, and fire extinguisher are located. Know how to use them.
3.	Insist upon good housekeeping in your laboratory.
4.	Check for insects and rodents. Keep all areas clean.
5.	Secure all gas cylinders.
6.	Wear laboratory coats and other appropriate protective clothing while performing laboratory activities. Feet and legs should be covered; sandals and open-toed shoes should not be worn in laboratories. Wear appropriate gloves while handling infectious or toxic materials and animals. Do not wear lab coats. Gloves or other personal protective equipment outside the laboratory.
7.	Use a biological safety cabinet for handling infectious materials or materials requiring protection from contamination and a fume hood for toxic materials; mixed hazards need to be evaluated case by case.
8.	Fume hoods should be used for laboratory activities that could result in chemical explosions or fires, for experiments involving toxic, hazardous or carcinogenic compounds, and use of strong acids and bases. Biological safety cabinets should not be used for this kind of work. Fume hoods are workstations, not storage cabinets. Vented storage areas may be located under the fume hood work area. However, these are not for storage of flammables.
9.	Respect chemicals and radionuclides. Know their hazards and follow appropriate safety precautions. Chemical and radioactive waste must not be poured down the drain.
10.	All equipment must be documented to be free of chemical, biological, and radiological contamination before repair work is done or before moving equipment for storage or elsewhere.
11.	Never mouth pipette anything. Use mechanical pipetting devices only!
12.	Close laboratory doors while experiments are in progress. Restrict access to laboratory.
13.	Put liquid traps and in-line vacuum filters on all vacuum lines.
14.	Minimize or contain all aerosol-producing activities, large-volume work, concentrated solutions or cultures. These activities include centrifugation (use safety cups), vortex missing (stopper tube), blending (use metal safety blender), sonication, grinding, opening containers of infectious material, inoculating culture flasks, inoculating animals, harvesting infectious materials from cultures or animals, and weighing or reconstructing toxic powders, etc.
15.	Place biological safety cabinets in low-traffic areas and minimize activities that disrupt air flow in or around cabinet.
16.	Decontaminate all work surfaces daily, and decontaminate all spills immediately.
17.	Decontaminate (by autoclaving or chemical disinffection) all biologically contaminated materials – glassware, animal cages, laboratory equipment, etc. – before washing, reuse or disposal. Discard materials via proper waste stream.
18.	Be careful with needles and syringes. Use only when alternative methods are not feasible.
19.	Syringes, needles, Pasteur pipettes, etc, should be placed in rigid, leak-proof containers (Sharps Safe) and discarded following the waste rules.
20.	Broken glassware and disposable pipettes (after decontamination) should be placed in a “Disposable Labware and Broken Glass Box” and discarded following the waste rules.
21.	Place contaminated biological materials in covered, leak-proof containers before removing them from the laboratory.
22.	Wash your hands after handling chemicals, infectious materials, animals, after removing gloves and before leaving the laboratory.

Trogene Horse's Hometown

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Team Members	Project	Parts	Human Pratics	Modeling	Safety	Notebook	Attributions

Lost the way? Use it to help you if you're lost.

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 <br>
 <p class="qbody"><span class="qnum"><font size="3" face="Verdana" color="#90B849"><strong>a)</strong></font></span><strong><font size="3" face="Verdana" color="#333">What is the Safety Level of your lab? (Use the <a href="https://2014.igem.org/Safety_Hub#rgbsl" target="_blank">WHO numbering system</a>, in which Level 4 is used for the most dangerous organisms.)</font></strong><font size="3" face="Verdana" color="#333"></font></p>
-<div class="wrapper" style="background-color: white;"><font size="3" face="Verdana" color="#333"><ul style="margin-left: 1em;"><li><a href="https://2014.igem.org/Safety_Hub#safety_levels" target="_blank">Level 2</a> (moderate risk, ~= WHO BSL 2)</li></ul></font></div>
+<div class="wrapper" style="background-color: white;"><font size="3" face="Verdana" color="#333"><ul style="margin-left: 1em;"><li><a href="https://2014.igem.org/Safety_Hub#safety_levels" target="_blank">Level 2</a> (moderate risk, &#8780; WHO BSL 2)</li></ul></font></div>
 <br>
 <p class="qbody"><span class="qnum"><font size="3" face="Verdana" color="#90B849"><strong>b)</strong></font></span><strong><font size="3" face="Verdana" color="#333">According to the WHO Biosafety Manual, what is the BioSafety Level rating of your lab? </font></strong></p>