Making a long term stock of bacteria

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Whenever you successfully transform a bacterial culture with a plasmid or whenever you obtain a new bacterial strain you will want to make a long term stock of that bacteria.  Bacteria can be stored for months and years if they are stored at -80C and in a high percentage of glycerol.   
Whenever you successfully transform a bacterial culture with a plasmid or whenever you obtain a new bacterial strain you will want to make a long term stock of that bacteria.  Bacteria can be stored for months and years if they are stored at -80C and in a high percentage of glycerol.   
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==Lab Protocols==
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==Materials==
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*40% glycerol solution
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* Day/overnight culture
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*Cryogenic vials/1.5mL microfuge tube
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==Method==
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*Add 1 ml of 40% glycerol in H<sub>2</sub>O to a cryogenic vial.
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*Add 1 ml sample from the culture of bacteria to be stored.
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*Gently vortex the cryogenic vial to ensure the culture and glycerol is well-mixed.
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**Alternatively, pipet to mix.
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*Use a tough spot to put the name of the strain or some useful identifier on the top of the vial.
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*On the side of the vial list all relevant information - part, vector, strain, date, researcher, etc.
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*Store in a freezer box in a -80C freezer.  Remember to record where the vial is stored for fast retrieval later.
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==Notes==
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*While it is possible to make a long term stock from cells in stationary phase, ideally your culture should be in logarithmic growth phase.
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==Specific Protocols==
*[[Endy:Making a long term stock of bacteria]]
*[[Endy:Making a long term stock of bacteria]]
*[[Endy:Retrieving a Registry glycerol]]
*[[Endy:Retrieving a Registry glycerol]]
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*[[-80_Glycerol_Stocks]]
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*[[Alm:Glycerol_stocks]]
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*[[McClean:E._coli_Glycerol_Stocks]]
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*[[Cfrench:glycerol]]
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*[[Smolke:Protocols/Freezer_stocks]]
==Notes==
==Notes==

Revision as of 04:34, 28 June 2012

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Contents

Introduction

Whenever you successfully transform a bacterial culture with a plasmid or whenever you obtain a new bacterial strain you will want to make a long term stock of that bacteria. Bacteria can be stored for months and years if they are stored at -80C and in a high percentage of glycerol.

Materials

  • 40% glycerol solution
  • Day/overnight culture
  • Cryogenic vials/1.5mL microfuge tube

Method

  • Add 1 ml of 40% glycerol in H2O to a cryogenic vial.
  • Add 1 ml sample from the culture of bacteria to be stored.
  • Gently vortex the cryogenic vial to ensure the culture and glycerol is well-mixed.
    • Alternatively, pipet to mix.
  • Use a tough spot to put the name of the strain or some useful identifier on the top of the vial.
  • On the side of the vial list all relevant information - part, vector, strain, date, researcher, etc.
  • Store in a freezer box in a -80C freezer. Remember to record where the vial is stored for fast retrieval later.

Notes

  • While it is possible to make a long term stock from cells in stationary phase, ideally your culture should be in logarithmic growth phase.


Specific Protocols

Notes

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