2.6: Appendix IV- Preparing Glycerol Stocks
- Page ID
- 67605
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Making a Glycerol Stock
ONLY if you have confirmed that your NEW composite part (newly built plasmid) is CORRECT. This means that after conducting a Zymo Miniprep and restriction digest, the new plasmid now contains the parts that you have ligated together.
How to make a glycerol stock?
- Using sterile technique, aliquot 500µL of 80% glycerol into a cryovial (This may or may not have been done already, ask your instructor)
- Label the cryovial clearly on top or on the side with the part number, the antibiotic resistance (amp, chlor, tet, kan), as well as any other labeling standard your lab uses (LABELING IS VERY IMPORTANT!!)
- Vortex your culture
- Carefully add about 500uL (0.5mL) of your culture to the cryovial. Vortex and place the vial on ice.
Store the vial in a -80 freezer that has been designated for long term storage.