Rickus Lab Protocols

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Contents

Lab Waste

General Lab Waste Procedures Created by: J.L. Rickus Created date: 4/19/2005, updated 6/9/2006, updated 11/26/2007

All waste generated by the lab is to be disposed of in proper containers. Waste is sorted by type. There are trash containers around the lab with labels that indicate their contents. Different waste types have different disposal procedures.

DRY WASTE

  • Lab Glass: All glass waste (disposable glass vials, nonbioglass pipets, microscope slides, empty chemical vials) that has not been exposed to hazardous chemicals should be placed in a lined box labeled as "Sharps". All tubes and vials need to be empty, containing no liquids or chemicals. When the box is full, seal the box, label it as glass sharps, and place it with the normal trash to be picked up for disposal by the janitor.
  • Plastic Lab Waste: Plastic pipettes, empty plastic tubes, empty eppendorf tubes should go into a separate ridid container. There should be NO liquids in any of the tubes. When the container is full, seal it up and label it “Sharps”. This can go out with the normal trash.

LIQUID WASTE

  • Hazardous Chemicals: Familiarize yourself with the Chemical Hygeine Plan! This document is the authority.
  • Alcohols
  • Acids and Bases

never put anything in the sink unless you are sure you can safely and legally do so!

BIOLOGICAL WASTE See the PURDUE REM page on Biological Waste as a reference

  • Solid Cell Culture Waste:
    • Glass Pipets: Place in plastic tub under the cell culture hood. When this fills up, transfer pipets to a lined cardboard box. Seal the box, label the box as “Glass Sharps” and place with the standard trash.
    • Cell culture plates, plastic pipets, tubes that contained cells and any other biohazard-look-alike-items should be placed in the special container labeled “cell culture waste”. This can contains an autoclave bag. When it is full, seal it with auto clave tape, autoclave it, box it and seal it in a box, tape a white Bio-Materials and Treatment Certification form on it. Call REM to pick it up. No fluids in this bag! Aspirate all fluids from the waste before putting it in the container.
    • Other: Kimwipes, gloves, paper towels can go into a normal trash container.
  • Other: Most other dry waste (other than chemicals) that we generate can go into the normal trash. If you are not sure, please ask.
  • Liquid Cell Culture Waste:
    • all liquid waste should get sucked into the vacuum flask containing sufficient disinfectant such as betadine or bleach. Once properly disinfected this liquid can go down the laboratory drain.

Special Notes on Chemical Handling

  1. APTMS & APTES. This is a preventative note to everyone who works in the biopolymer "chemistry" area. Many of us are involved in materials work or chemical functionalization involving silane-type chemicals. If one of these chemicals happens to spill or is dripped onto the threading of the bottle - it can and will fuse the bottle in a permanently closed position, making the remaining chemical unobtainable and therefore wasted. In particular, this refers to highly reactive conjugative silanes, such as the aminopropyltrimethoxysilane (APTMS) and aminopropyltriethoxysilane (APTES).

These bottles should please be kept upright during storage (as with any liquid) and if a spill occurs or if the chemical is dripped onto the cap or bottle closure threading - it needs to be wiped cleaned with methanol or ethanol (for the threading) or a mixture of ethanol and water for the bench. Additionally, the bottle in question should be checked after a day to ensure that it is still usable, and if not, a new stock ordered. Accidents do happen, but please try to remember that we sometimes all use chemicals from the same stock, and if the stock is depleted or otherwise inaccessible, it can slow all of our research.

Cell Culture

[|Liquid Nitrogen Inventory]

Please note changes as they occur.

  1. Preparation to work in cell culture Please be sure to wash your hands and arms thoroughly, and wear clean gloves sprayed with ethanol prior to reaching into the cell culture incubator.
  2. Monthly Check of Cultures Near the first week of each new month, split off a small part of your culture to be tested for contamination. This small aliquot should be plated as usual and assayed after ~3-4 days in culture without antibiotics.
  3. Autoclaving the Incubator Trays Everything inside of the incubators, including the trays, are autoclavable. Sanyo's recommendations for cleaning.Media:Example.ogg

Biochemistry

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