Difference between revisions of "Team:Toronto/Experiment-Glycerol Stock Protocol (Bacteria)"

 
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Latest revision as of 06:58, 19 October 2016

Glycerol Stock Protocol (Bacteria) · Benchling

Glycerol Stock Protocol (Bacteria)

Introduction

Get started by giving your protocol a name and editing this introduction.

Materials

  • Reagent
    • Glycerol
    • Overnight desired bacteria culture
    • Appropriate antibiotic
  • Equipment
    • 2mL microcentrifuge tube
    • Sterile pipette tips
    • 10mL Falcon tube
    • LB plate

Procedure

  • How-to
  1. Add 500μL 40% glycerol to a 2mL microcentrifuge tube.
  1. Add 500μL overnight culture of desired bacteria.
  1. Transfer the glycerol stock to the -80°C freezer. (AZ: If possible, its better to flash freeze with liquid nitrogen and then put into -80 freezer)
  1. To recover bacteria, open the tube and use a sterile pipette tip (bring the pipette tip box to the freezer, don't put the tip on in the lab and walk there) to scrape some of the frozen bacteria off the top. Transfer right there to a 10mL Falcon tube.
  • ␥ Do not let the glycerol stock unthaw.
  • ␥ Do not bring it back to the lab to transfer with the pipette tip – thawing the glycerol stock will decrease the efficiency of any downstream applications for the bacteria that will remain in storage after you're done.
  1. Start an overnight culture with the appropriate antibiotic (antibiotic added in 1:1000 dilution – i.e. 10μL antibiotic in 10mL LB).
  1. The next morning, streak onto an LB plate, and continue as necessary.
  • Changelog
  • 1. Created 15/5/2016



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