Jan 07, 2020
Preparation of Frozen Bacteria Stock
- 1University of Arizona
- Yoon Lab
Protocol Citation: Kenneth Schackart 2020. Preparation of Frozen Bacteria Stock. protocols.io https://dx.doi.org/10.17504/protocols.io.ba3bigin
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: January 07, 2020
Last Modified: January 07, 2020
Protocol Integer ID: 31555
Abstract
This protocol details how to prepare bacteria culture for deep freezing. Protocol adapted from the protocol provided as an external link.
Reference:
Vitko N.P. and Richardson A.R. 2013. Laboratory Maintenance of Methicillin-Resistant Staphylococcus aureus (MRSA). Curr Protoc Microbiology. Basic Protocol 4: Preparation of S. aureus Frozen Stocks.
Guidelines
Gloves must be worn at all times. Observe aseptic technique to reduce the risk of contamination.
Materials
MATERIALS
DMSOSigma AldrichCatalog #D1435
- Crytotube(s)
- Overnight bacteria culture in broth
- Dimethyl sulfoxide (DMSO)
Before start
Suggested: sterilize crytoube in biosafety cabinet UV.
Prepare Stocks
Prepare Stocks
Add 150 µL sterile DMSO to sterile cryotube.
Add 850 µL overnight culture to cryotube containing.
Vortex or invert repeatedly to mix.
Store cryotube in deep freezer (-40 °C to -80 °C )
Update Inventory
Update Inventory
Go to the frozen storage inventory and add at a minimum the following information:
- Storage location
- Your name
- Date
- Organism identification