BRL (Biofuels Research Lab) Protocols A training aid for all new and experienced users of the Biofuels Research Lab Ed Evans Research Support Specialist Updated

General Usage Guidelines Safety Glasses MUST be worn in the BRL labs at all times – Corrective eyeglasses with safety lenses may be worn unless specific PPE (Personal protective equipment) is required All users MUST take EH&S safety courses (Laboratory safety, Chemical Waste disposal, Fume hood safety, How to read an MSDS, Fire Safety, and Cleaning small lab spills) before using the BRL – More courses are required if using specialized equipment (centrifuge safety, cryogen safety, etc.)  All BRL users must be trained via a BRL Specialist prior to using ANY equipment Chemical MSDS sheets are located in the main hallway outside B77 and available through chemical inventory on BRL Network (Enterprise) BRL users must be activated in system via Ed Evans (ece22) or Sue Fredenburg (saf3) prior to gaining access BRL users may not use USB drives on any lab computer Personal folders are located on BRL network (Enterprise) and are used for data storage Only computers in the BRL office can be used to transfer data out of the lab

BRL Server/Network (Enterprise) Sophisticated and secure network to protect computers associated with measured analytical platforms Provides secure back-up for large quantities of data Controls user access to data and analytical platforms File systems are organized with different levels of access and security Can be connected from anywhere via VPN

Fume Hood General Knowledge Keep all materials inside the hood at least six inches from the sash opening. When not working in the hood, close the sash. Be sure that nothing blocks the airflow through the baffles Never allow your head to enter the plane of the hood opening. Elevate large equipment at least two inches off the base of the hood interior Do not permanently store any chemicals inside the hood. Promptly report any hood that is not functioning properly to your supervisor When using extremely hazardous chemicals, understand your laboratory’s action plan in case an emergency What is wrong with this picture?

Centrifuges (Bench-top and Large Volume) All centrifuge users must be trained on centrifuge usage and take the EH&S centrifuge training course All users must sign in with the log sheet User name, , date, and rotor type Carbon Fiber Rotors must be stored properly within specific canvas bags and not placed on benches without protection Samples must be balanced when placed in rotors Once finished with centrifuge, the lids are to be open and the unit turned off Any spills must be cleaned up by user All containers must be cleaned and sterilized

Ensure that centrifuge bowls and tubes are dry Ensure that the spindle is clean Use matched sets of tubes, buckets and other equipment Inspect tubes or containers for cracks or flaws before using them Avoid overfilling tubes or other containers Ensure that the rotor is properly seated on the drive shaft Make sure that the centrifuge is operating normally before leaving the area Do not exceed the rotor’s maximum run speed – Safety feature of BRL centrifuges Report all accidents and malfunctions to supervisor immediately Centrifuges (Bench-top and Large Volume)

Autoclave Usage Bench-top or Large Volume Personal Protective Equipment (PPE) must be worn when using autoclaves – Autoclave gloves are stored in B77 (Research Support Lab) across from the large volume centrifuges All users must be trained by Ed Evans (ece22) – All users must sign receipt after run Ensure that the autoclave/sterilizer door is closed and locked before beginning the cycle Do not remove items from an autoclave until they have cooled Use forceps or other tools to remove instruments and other items from baskets and autoclaves Items must be loaded with autoclavable containment bins

-80°C Freezers All users must be trained and given a specific storage rack All users must take EH&S course (Cryogen safety) Avoid skin and eye contact Cryogenic gloves must be worn whenever handling items in -80°C Freezer Never store a cryogen in a sealed, airtight container at a temperature above the boiling point of the cryogen First Aid In case of exposure to cryogens or dry ice, remove any clothing that is not frozen to the skin. Do NOT rub frozen body parts because tissue damage may result. Obtain medical assistance as soon as possible. Place the affected part of the body in a warm water bath To report an emergency, dial 911

Material Handling Lab Must be trained on equipment prior to use Due to the nature of equipment in this lab, PPE’s are required Ear plugs or Muffs must be worn Face shield and dust mask must be worn Pinch points

Chemicals Need to know Chemicals are stored within the Support Lab, alphabetically unless specific storage needs are required All chemical MSDS sheets must be stored in MSDS folder located in main hallway near telephone, organized via room and alphabetically Any chemical brought into the BRL must be accompanied by the MSDS form and filed properly (and added to online chemical inventory) The identity of the chemical and appropriate hazard warnings must be shown on the label The hazard label message must be legible, permanently displayed and written in English

Hazardous Chemical Waste Chemical waste must be stored in the same lab in which it is generated – A “Hazardous Waste Satellite Accumulation Area” sign should be posted at the area where the waste is generated and storedHazardous Waste Satellite Accumulation Area Complete a green Hazardous waste tag. Peel and stick the bottom copy to the waste container Tags are stored in B77 (Research Support Lab) across from the large volume centrifuges All waste must be marked as Hazardous Waste Contact Ed Evans (ece22) for waste removal from BRL

Hazardous Chemical Waste Continued All generators must attend Chemical Waste Disposal Training Waste containers must be in good condition, no dents, cracks etc., and lids intact and functional. Wastes must be chemically compatible with the container. Containers must be closed except when adding or removing waste. Do not leave a funnel in the bottle. Store hazardous waste chemicals in secondary containment whenever possible. Plastic bins offer the best protection against spills. Waste containers must be under the control of the operator of the process that generates the waste.

Chemical Inventory The Chemical inventory can be located from any computer connected to Enterprise (or via VPN) – Enterprise → Ten Forward → Chem Inventory 2012 Separated via dry and wet chemicals Access to pdf of MSDS by clicking on chemical Indicates supplier, grade, location, catalogue #, CAS #, and Approximate volume/weight Must be updated whenever new chemicals are brought into lab or consumed

Contaminated Waste Disposal Material Contaminated with Biohazards Materials 1 Contaminated with Non-Biohazards Materials 2 Syringes (no needle at any time)Sharps Disposal Container/ RMWPuncture Resistant Container/ Regular Trash Syringes with attached needles (hypodermic, HPLC, GC) Needles Razor blades Scalpels Blood vials Sharps Disposal Container/ RMW Pasteur or serological pipettes Micropipette tips Glass slides and cover slips Glass tubes and other glassware Broken plastic ware Sharps Disposal Container/ RMW 3 Puncture Resistant Container/ Regular Trash Plastic Petri dishes Plastic tubes, flasks, plates Gloves Bench paper and towels Disposable gowns Red Biohazard Bag/ RMWRegular Trash Cell culture wastes Body fluids Treat with Disinfectant or Autoclave/ Drain dispose (Sanitary Sewer only) Animal carcasses Tissues Bedding Red Biohazard Bag/ Incineration 4 Black Bag/ Incineration 4 Ethidium bromide- contaminated lab waste Gels Red Biohazard Bag or Sharps Disposal Container/ RMW 5 Per EH&S Guidelines 6 Mixed WastesContact EH&S Cornell EH&S Biological Waste Disposal Guide

Regulated Medical Waste (RMW) Disposal All regulated waste must be segregated into Red Biohazard Bags, Incinerator Cartons, or Sharps containers. The RMW tags are no longer needed, however all waste must still be labeled and attach to EACH container. – Cartons must be sealed when waste reaches approximately 1 inch from the top – Tags can be obtained in the Research Support Lab (B77) in a drawer across from the large volume centrifuges Name: Section/Lab: Building: Riley-Robb Hall BRL Phone number: Ensure bags and cartons are sealed with zip ties and/or packing tape. Ensure sharps container lid is secure. Gloves must be worn at all times. Place bags, cartons, or sharps containers on cart to move. Sharps containers or small containers may be carried by hand. Transport containers to Dept. Autoclave room (room across from elevator in basement) Place waste in RMW container (see picture) or place incinerator cartons on floor to the side of the removal bin. EH&S Contact for RMW Removal: Bill Leonard ( ) or Biofuels Research Labs Contact: Ed Evans ( ) or Contact EH&S ( ) in case of any large spills or release of material.

Final General Policies Anything that is borrowed from any lab must stay within the BRL and be returned to the place it was borrowed by the end of that workday (unless it is being used for an extended experiment and others are aware of the item’s location) Many disposable items in the BRL are for general use and can be found in the support lab or on the racks behind the growth chambers. Other non-disposable items are found in the other labs and have been purchased for specialized projects. These items require permission from users to borrow especially if they come from a designated student’s bench or drawer. Otherwise, the item needs to be purchased by the user Pipettes are BRL property unless purchased by an outside user. Once a user is done with an area, the area must be cleaned by the user. This applies not only to workspaces but balances, glassware, and equipment. All liquids and chemicals must be labeled, including water and waste containers. Please see Ed Evans or Cornell EH&S training for any questions. Contaminated Gloves are not to be worn outside or the lab in which you are working – Do not touch door handles with contaminated gloves

BRL Contact Info Larry Walker, Ph.D.: Facility Principle Investigator or Edward Evans, MS: Facility Research Support Specialist or Tamara Payne: BRL Financial Representative or

EH&S Training and Contact Info  To report an emergency, dial 911 Contact EH&S: Ask EH&S: The online question and answer system is the best way for the Cornell community to ask Environmental Health and Safety questions to EH&S staff:  EH&S Training Website: 