Altman:Protocols/MB Protocols/Glycerol stock: Difference between revisions
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1. Prepare a glass test tube with 5 mL of TB+antibiotic | 1. Prepare a glass test tube with 5 mL of TB+antibiotic | ||
2. Scrape a chunk of the glycerol stock with a sterile loop | 2. Scrape a chunk of the glycerol stock with a sterile loop, and place in the test tube | ||
3. Shake or spin at 37C for no more than 18 hours | 3. Shake or spin at 37C for no more than 18 hours | ||
Revision as of 19:52, 14 September 2014
Starting a bacterial culture from a gycerol stock
- Prepare TB (terrific broth) beforehand. TB can be used for a few weeks, and then must be discarded.
1. Prepare a glass test tube with 5 mL of TB+antibiotic
2. Scrape a chunk of the glycerol stock with a sterile loop, and place in the test tube
3. Shake or spin at 37C for no more than 18 hours
- Tubes should be cloudy with cells. Be sure to do a control, which should appear clear at the end of 18 hours
4. Move the 5 mL of culture to a 15 mL conical and spin in a centrifuge at 4000 rpm for 15 minutes
5. Remove the supernatant
6. If you are not going to do the Miniprep that day, place the cells in a -20C freezer
