Lyophilizer usage

How to Use the Lyophilizer to freeze dry buffers:

Aqueous buffers ONLY

NO ORGANIC SOLVENTS (ie. ethanol)

1. Check if condenser is dry inside, if not mop up water
2. Check if valves are closed (oriented with bevel up)
3. Turn on vacuum pump; check for bubbles (should not have water inside)
4. Turn on lyophilizer (switch on right side)
5. Press auto button (will refrigerate to -50 and then automatically turn on vacuum). Wait until temp and vacuum are stable (stable green light)

a. The lyophilizer is ready when temp and vacuum are stable (w green light) AND when pressure is < 0.1 mBar (ideally 0.04 mBar) b. If pressure won’t go down, there could be a leak

6. Meanwhile, freeze your buffer in dry ice + EtOH bath or LN2

a. Use a container that is at least 5x volume of your buffer b. Make sure buffer is very frozen (want rate of sublimation > rate of melting)

7. Fold up a kimwipe and place over open conical. Secure with a rubber band. Quickly to avoid melting
8. Place into bell jar containing paper towel (to soak up condensation). Choose correct size rubber lid (check that it contains a filter). Secure into a rubber port on the lyophilizer and turn the corresponding valve to begin vacuuming contents of jar. Pressure will rise so make sure that it goes back down.
9. Check in a few hours to make sure dehydration is occurring. Should ideally run 24h but O/N may be sufficient
10. If you want to add a sample to the jar, turn valve to closed first and then add to jar.
11. Turning off: press vacuum button, refrigeration button, turn off lyophilizer, turn off vacuum pump
12. Store samples in freezer to prevent back rehydration