3.07. Preparing Glycerol Stocks

Glycerol stocks are used for long-term storage of plasmids. This allows you to easily obtain the desired bacterial strain with the desired plasmid without having to transform more cells.

1. Prepare five 1 ml cultures of desired bacteria.
2. Allow cultures to grow overnight.
3. Pipette 15% desired volume of glycerol into tubes.
4. Pipette 85% desired volume of bacterial culture into tube.
5. Tighten the caps of the tubes.
6. Shake/vortex tubes.
7. Pour liquid nitrogen into a container.
8. Use tweezers to submerge the first tube until frozen.
9. Store the tube in -80°C.
10. Repeat steps 7 and 8 with the remaining tubes.

https://www.addgene.org/recipient-instructions/myplasmid/