Team:Washington/Protocols/Glycerol Stocks

From 2011.igem.org

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=Glycerol Stocks=
=Glycerol Stocks=
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''For the long-term storage of microbial cultures''
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For the long-term storage of microbial cultures
''Make sure that you do not contaminate glycerol stocks''
''Make sure that you do not contaminate glycerol stocks''
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1. Grow overnight culture (1 mL) of your cells in selective media
1. Grow overnight culture (1 mL) of your cells in selective media
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2. Mix 750 uL of your culture with 750 uL of 40% glycerol.  Immediately place at -80 C (glycerol is toxic to cells and prolonged exposure to it kills them at RT).
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2. Mix 500 uL of your culture with 500 uL of 50% glycerol.  Immediately place at -80 C (high concentration of glycerol is toxic to cells and prolonged exposure to it kills them at RT).
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If you need to take from the stocks, do not thaw - just scrape ice off of the top with a sterile loop and smear across a plate.
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If you need to take from the stocks, do not thaw - scrape ice off of the top with a sterile loop and streak across plate.

Latest revision as of 01:38, 14 September 2011


Glycerol Stocks

For the long-term storage of microbial cultures

Make sure that you do not contaminate glycerol stocks

1. Grow overnight culture (1 mL) of your cells in selective media

2. Mix 500 uL of your culture with 500 uL of 50% glycerol. Immediately place at -80 C (high concentration of glycerol is toxic to cells and prolonged exposure to it kills them at RT).

If you need to take from the stocks, do not thaw - scrape ice off of the top with a sterile loop and streak across plate.

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