Shreffler:Aseptic Technique
General Guidelines
- Protect eyes, mucous membranes, open cuts and wounds from contact with biohazardous material.
- Use personal protective equipment as necessary (i.e. gloves, goggles, mask, other protective clothing).
- Carry out all culture operations under a laminar flow hood.
- Disinfect all surfaces with a disinfectant solution prior to use.
- Swab down the working surface liberally with 70% ethanol solution.
- Periodically spread 70% ethanol solution over the exterior of gloves to minimize contamination.
- Replace gloves if torn.
- In case of any spill, spread 70% ethanol solution and swab immediately with non-linting wipes.
- Discard gloves after use. Do not wear them when entering any other lab area.
- Bring into the work area only those items needed for a particular procedure.
- Leave a wide clear space in the center of the hood (not just the front edge) to work on.
- To prevent blockage of proper air flow and to minimize turbulence, do not clutter the work area.
- Swab all glassware (medium bottles, beakers, etc.) with 70% ethanol solution before placing them inside the hood.
- Arrange work area in order to have easy access to all items. Do not reach over one item to retrieve another (especially over an open bottle or flask).
- Use sterile wrapped pipettes and discard them into a biohazard waste container after use. Check that the pipette wrapping is not broken or damaged, to confirm sterility.
- Inspect the vessels to be used:
- T-flask
Back to Lab Protocols • Check that the wrapping of the sterile pipet is not broken or damaged. • Inspect the vessels to be used: • T-flask - Must be free from visible contamination or breakage, or lack container identification. The plastic covering the flask must be intact. • Bottles - Check for cracks, expiration dates. • Spinner flasks - Check for cracks, expiration dates, and proper assembly. • Discard any biohazard or contaminated material immediately.. • Never perform mouth pipetting. Pipettor must be used. • When handling sterile containers with caps or lids, place the cap on its side if it must be laid on the work surface. • Make sure not to touch the tip of the pipette to the rim of any flask or sterile bottle. • Clean the work area when finished by wiping with 70% alcohol. 18.
