Sterilization

Sterilization kdorfman Tue, 06/16/2009 - 17:50

Sterilize anything that cells might grow in.

Autoclave glassware, tips, and microfuge tubes.

Autoclave most media and salt solutions.

Do not sterilize by autoclave items containing:

  • detergents (e.g., SDS) - they can boil over

  • heat sensitive ingredients (e.g., vitamins, hormones, antibiotics, proteins)

  • sugar in growth medium (the sugars and amino acids may react together, reducing the concentration of both)

  • HEPES

  • DTT (dithiothreitol)

  • Beta mercaptoethanol

  • corrosives (e.g. acids, bases, phenol)

  • solvents or volatiles (e.g. ethanol, methanol, chloroform, acetone, formaldehyde, formalin or glutaraldehyde)

  • chlorine (e.g., bleach)

  • anything radioactive

Filter sterilize any liquid that must be sterile, and that you cannot autoclave.

biohazard_waste_only.docx

Autoclave

Autoclave kdorfman Tue, 06/16/2009 - 20:25

autoclave manual

Always put a piece of autoclave tape on the item or container so you can tell if it was exposed to the steam.

If the green light is on, press the red reset button.

  • Biohazard Waste: See here: https://wahoo.nsm.umass.edu/content/biohazard-waste

  • Tips: Load racks into boxes wearing gloves (this caution is primarily for RNA work, as most people's skin has RNase on it).
    15 minute sterilization; 40 min drying time; open autoclave CAUTION - HOT! to let steam escape.
    If there is too much condensation inside the boxes, put them in the oven at ~60C for ~an hour.

  • Microtubes: Put into 600 mL plastic jars, screw cover on loosely, so the steam will penetrate. Same time as for tips.

  • Liquids: Larger volumes require longer sterilizing times. Use this table:

Largest volume (mL) Minimum time (min)
75 25
250 30
500 40
1000 45
1500 50
2000 55
>2000 55 + 10 per L
  • Monthly spore test EH&S recommends Fisher 12-001-1 population 10^5 Prospore Bacillus stearothermophilus
To sterilize mL of liquid medium
autoclave for: minutes

Minutes = 8.3853 * volume^0.2449

autoclave_log.docx
autoclavemanual.pdf

Monthly Maintenance

Monthly Maintenance kdorfman Fri, 09/09/2016 - 15:45

  • Turn generator switch off

  • Let cool to ~5 lb pressure

  • Turn master switch off

  • Open valve

  • Let tank drain

  • Turn generator switch on

  • Close valve

  • Let it fill

  • Turn master switch on

Autoclave bags

Autoclave bags bcrcstaff Thu, 02/14/2019 - 13:45
Container dimensions (in) bag size
Rubbermaid 16" x 10" 24" x 36" Fisher 01-8143
Round diam = 18" 38" x 48" VWR 14220-044
Square 16" x 14" 38" x 48" VWR 14220-044