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Laboratory Rules Access to the Laboratory ■ Only the Chiefs of Laboratories Shall Have the Key of Their Assigned Laboratory

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Laboratory Rules Access to the Laboratory ■ Only the Chiefs of Laboratories Shall Have the Key of Their Assigned Laboratory Laboratory Rules Access to the Laboratory ■ Only the chiefs of laboratories shall have the key of their assigned laboratory. ■ The presence of unauthorized personnel in the laboratory is strictly prohibited. ■ Before starting activities in the laboratory, users must attend the Work Induction Course in the Laboratory. Use of Equipment and Material in the Laboratory ■ For each laboratory equipment there is a log of use, in which each user must write down the following information: ■ Name of the user ■ Date ■ Type of sample ■ Analysis to be performed ■ State of the equipment ■ Required time of use ■ In case of any problem with its use or damage, report it immediately to the academic technicians and the Head of the Laboratory. ■ The scales must be cleaned after use ■ The glassware material should be washed at the end of its use ■ The user must clean the areas of the laboratory that he has used at the end of his work. ■ For no reason should material and / or glassware be left for more than two days, outside of their assigned drawers. Use of Reagents ■ Under no circumstance should the remaining reagents be returned to their original containers, since contamination of the entire lot is possible. ■ Every reagent and solutions must be tightly closed and labeled properly (reagent name, preparation date, user name). ■ Reagent solutions should be stored in bottles or in suitable containers, for a period not exceeding three months. It is not allowed to store them in laboratory materials such as beakers, flasks, test tubes, etc. Personal Safety ■ It is prohibited to enter the laboratories without the minimum safety equipment: buttoned gown, long pants, closed and low shoes, as well as protective glasses. ■ It is prohibited to eat, drink and store food in refrigerators, drawers or drawers in laboratories. The use of earphones or cell phones in the laboratory is not allowed. ■ No reagent should be touched, smelled or tasted. ■ Hazardous or toxic reagents and materials must be handled in the specific areas designated for them. ■ It is forbidden to leave the laboratory area wearing gloves and / or gown as well as answering the landline, touch the keyboards of computers or equipment, with the gloves on. Waste Management ■ Chemical waste will be classified and disposed of according to its nature. Users should consult the safety data sheets to know what type of waste is being generated. ■ Hazardous waste containers will be properly labeled and stored in a specific place until they are collected by the corresponding department (DEPARTMENT. ENVIRONMENT AND SAFETY FCQ). ■ Be sure to empty the waste in the corresponding container. Before using the container, check the level of the waste, -so as not to saturate the container, do not fill more than 4/5 parts of the container. Be sure to cover the container once the waste is poured. ■ Solid waste other than chemicals can be placed in the trash cans. ■ Do not put needles or sharp objects in regular trash containers. These must be disposed of in special containers. Accident Management ■ The chiefs of the laboratories, and the administrative staff must be immediately notified in the case of any accident. Local emergency telephone numbers located in every lab should be called if needed, found at the end of this regulation and attached to a visible place in the laboratory. ■ In case of fire: Only if you just start the fire try to extinguish it with the appropriate extinguisher. If the fire is of such magnitude that it is no longer possible to control with the fire extinguisher, call the laboratory managers. ■ If reagent contact occurs in the eyes, use the emergency eyewash immediately. Rinse for approximately five minutes and then seek medical help. ■ If corrosive liquids splash on skin or clothing, immediately rinse the affected areas thoroughly. If a large area is affected, use the emergency shower. If necessary, take off your contaminated clothes and then seek medical help. ■ For small cuts in the skin, it can be washed with soap and water and then put on a "band-aid". Do not work in the laboratory with any exposed wound. If the cut causes bleeding, stop it by pressing directly with a clean towel and seek help to go immediately to the emergency medical service. ■ For mild burns, apply the burn ointment on the affected area. Do not apply any other substance on the burn. For major burns, seek help to go immediately to a nearby emergency medical service. ■ If toxic gases are inhaled, move the victim to a ventilated place and ask for medical help. If required and if you are trained, administer artificial or cardiopulmonary respiration. ■ All accidents should be reported as soon as possible to the heads of the laboratory. ■ It is vital to know the location and the operating instructions of the following: Eyewash, Emergency showers, Fire extinguishers, First aid kit, Emergency stairs, Emergency numbers and Emergency exits. Legal Perspective Legal Perspective Mexico is part of two major international agreements: the Cartagena Protocol, which deals with biosafety issues, and the Nagoya-Kuala-Lumpur Protocol on Liability and Compensation. Both of them emerge from the Convention on Biological Diversity (CBD), which has other approaches for the conservation and sustainable use of biodiversity and the responsible use of genetic resources. The function of these different protocols is to guarantee an adequate level of protection concerning the transfer, management and use of living modified organisms (LMOs) of the current biotechnology application which may affect the responsible use of biological diversity and its conservation. These agreements promote different procedures to carry out safe handling, transfer and use of LMOs. This protocol proposes to examine different previous criteria depending on the situation. These previous criteria are based on the analysis of possible future effects of LMOs and evaluation of the extent of the damage if they are released before transfer. The Nagoya-Kuala Lumpur protocol is an international protocol which has the function of complementing the Cartagena protocol contributing to the conservation and sustainable use of biodiversity. The evaluation of other factors, such as risk to human health, based on responsible international standards and procedures, is critical. The objectives of this protocol are to seek to prevent, reduce, contain and avoid the possible damage caused by living modified organisms (LMOs). Protection Objectives Human: ■ Protecting the team’s safety in the work environment with an influential culture of prevention. ■ Ensuring that every member of the team involved with laboratory work is qualified to handle the material safely and are also prepared to handle possible risk scenarios related to the development of the project. ■ Protecting the community’s well-being by ensuring the isolation of the GMO and its genetic material, and also by avoiding the use of any pathogenic genes or species that may be harmful to humans, animals or plants. Environment: ■ Ensuring that the GMO and its genetic material remains contained within the limits of the laboratory setting. ■ Preventing the gene-leaking caused by horizontal gene transfer. ■ Avoiding the use of any pathogenic genes or the handling of species that may be harmful to humans, animals or plants. Laboratory: ​ ■ Ensuring the appropriate use of the laboratory equipment by following the standardized protocols ■ When possible, avoiding the use of harmful chemicals, such as Ethidium bromide, by replacing them with safer alternatives, such as GelRed; and when the use of a safer alternative is unfeasible, making sure that all of the preventive measures are being taken to prevent any harmful effects caused by the material Assessment of the Possible Risks Associated with the project Brief Description of the Project The iGEM FCB-UANL 2020 team project seeks to create, through the use of synthetic biology, an environmentally-friendly firefighting foam that can be a competitive alternative to the ones that are currently on the market. The foam will be produced based on three main components: ranaspumins, biofilm, and surfactin. Description of the Proposed Organism The final proposed GMOs consist of two organisms: an E. coli strain that integrates ​ genes from E. pustulosus and is able to produce 4 ranaspumins required to create a ​ stable foam structure.; and the other organism consists of an engineered B. subtilis ​ strain which does not sporulate and has an increased production of biofilm and surfactin. The parts used to construct the pathway to increase the production of surfactant agents in B. subtilis and to express Ranaspumins in E. coli were chosen because they do ​ ​ not have any significant safety concerns associated with them for their use in a laboratory. The parts used do not contain sequences that increase the virulence or any other risk factors in the recipient organism. Purpose of the GMO ■ General objective of the project: ■ Designing an expression cassette that enhances the expression of surfactin and biofilm in Bacillus subtilis ​ ■ Designing an expression cassette to express Ranaspumin 2, 3, 4 and 5 in Escherichia coli ​ ■ Short term objectives: ■ Using synthetic biology techniques to suppress the sporulation of Bacillus subtilis ■ Using synthetic biology techniques to synthetize the ranaspumin family of proteins (2 to 5) in Escherichia coli chassis ​ ​ ■ Using synthetic biology techniques to increase the production of surfactin in Bacillus subtilis ​ ■ Using synthetic biology techniques to maximize the production of biofilm in Bacillus subtilis ​ ■ Long term objectives: ■ To develop a competitive eco-friendly firefighting foam composed of natural surfactant proteins Identification of Potential Hazards of the GMO The recipient organisms: Escherichia coli K12 DH5α ​ Kingdom: Bacteria ​ Subkingdom: Negibacteria ​ Phylum: Proteobacteria ​ Class: Gammaproteobacteria ​ Order: Enterobacteriales ​ Family: Enterobacteriaceae ​ Genus: Escherichia ​ Species: Escherichia coli ​ Escherichia coli K12 DH5α is commercially available. The strains derived from E.coli ​ K12 are some of the most studied organisms in molecular biology, and they have been completely sequenced.
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