Team:Arizona State/Lab/Protocols/Cultures

From 2011.igem.org


Protocols: Cultures


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Liquid Culture:


  • Pick single colony from plate with pipet tip.
  • Drop tip in tube of ~7 mL LB media (with selected antibiotic if necessary).
  • Shake overnight at 37C.

Plating Cells:


From solution/liquid culture:

  • Apply 100-200 μl cell solution to LB agar plate containing appropriate antibiotic.
  • Place spreader in 95% ethanol, remove and sterilize in flame.
  • Using sterilized spreader, gently spread solution around plate.
  • Store upside down at 37C overnight to grow cells.
  • Store at 4C long-term.

Streak plating:

  • Place inoculation loop in 95% ethanol, remove and sterilize in flame.
  • Cool inoculation loop by pressing into a section of agar in used plate.
  • Pick single colony of cells from original plate.
  • Streak colony across new plate.
  • Store upside down at 37C overnight to grow cells.
  • Store at 4C long-term.

Glycerol Stock:


  • Prepare solution of 40% glycerol by volume.
  • Add 750 μl cell solution and 750 μl glycerol solution (or 1:1 mix) to Eppendorf tube.
  • Store at -80C.