Restocking Plasmids from Glycerol
Updated 5/24/2018 09:36 PM
Introduction
Materials
- Leucine Broth media
- Bacteria Glycerol Stock
- Culture Tubes
Procedure
Grow up more bacteria from stock.
Be as fast and as sterile as possible throughout this entire process.
- Prepare cell culture tubes with 4µL of LB media.
- Acquire the plasmid stock you want from the -80oC freezer.
- Scrape a glob of bacteria out of the stock using a 1000µL pipette tip. Stir into the culture tube, lightly fluff with pipette.
- Repeat step 3 for all of the plasmids you're restocking.
- Place culture tubes in 37oC shaker to grow up overnight.
Miniprep
- The next morning, remove the tubes from the shaker.
- Do a miniprep on each of the culture tubes to harvest the plasmids.
We have a frozen glycerol stock of bacteria with all of the plasmids we need! When our stocks of plasmid are running low, follow this procedure.