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Dehydrated Caenorhabditis elegans Stocks Are Resistant to Multiple Freeze-Thaw Cycles

Ultracold preservation is widely used for storage of genetic stocks of Caenorhabditis elegans. Current cryopreservation protocols are vulnerable to refrigeration failures, which can result in the loss of stock viability due to damage during re-freezing. Here we present a method for preserving worms...

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Autores principales: McClanahan, Patrick D., McCloskey, Richard J., Ng Tung Hing, Melanie, Raizen, David M., Fang-Yen, Christopher
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Genetics Society of America 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7718750/
https://www.ncbi.nlm.nih.gov/pubmed/33033066
http://dx.doi.org/10.1534/g3.120.401825
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author McClanahan, Patrick D.
McCloskey, Richard J.
Ng Tung Hing, Melanie
Raizen, David M.
Fang-Yen, Christopher
author_facet McClanahan, Patrick D.
McCloskey, Richard J.
Ng Tung Hing, Melanie
Raizen, David M.
Fang-Yen, Christopher
author_sort McClanahan, Patrick D.
collection PubMed
description Ultracold preservation is widely used for storage of genetic stocks of Caenorhabditis elegans. Current cryopreservation protocols are vulnerable to refrigeration failures, which can result in the loss of stock viability due to damage during re-freezing. Here we present a method for preserving worms in a dehydrated and frozen form that retains viability after multiple freeze-thaw cycles. After dehydration in the presence of trehalose or glycerol, C. elegans stocks can be frozen and thawed multiple times while maintaining viability. While both dauer and non-dauer larvae survive desiccation and freezing, the dauer defective mutant daf-16 does not survive desiccation. Our technique is useful for storing stocks in a manner robust to freezer failures, and potentially for shipping strains between laboratories.
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spelling pubmed-77187502020-12-17 Dehydrated Caenorhabditis elegans Stocks Are Resistant to Multiple Freeze-Thaw Cycles McClanahan, Patrick D. McCloskey, Richard J. Ng Tung Hing, Melanie Raizen, David M. Fang-Yen, Christopher G3 (Bethesda) Investigations Ultracold preservation is widely used for storage of genetic stocks of Caenorhabditis elegans. Current cryopreservation protocols are vulnerable to refrigeration failures, which can result in the loss of stock viability due to damage during re-freezing. Here we present a method for preserving worms in a dehydrated and frozen form that retains viability after multiple freeze-thaw cycles. After dehydration in the presence of trehalose or glycerol, C. elegans stocks can be frozen and thawed multiple times while maintaining viability. While both dauer and non-dauer larvae survive desiccation and freezing, the dauer defective mutant daf-16 does not survive desiccation. Our technique is useful for storing stocks in a manner robust to freezer failures, and potentially for shipping strains between laboratories. Genetics Society of America 2020-12-01 /pmc/articles/PMC7718750/ /pubmed/33033066 http://dx.doi.org/10.1534/g3.120.401825 Text en Copyright © 2020 McClanahan et al. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Investigations
McClanahan, Patrick D.
McCloskey, Richard J.
Ng Tung Hing, Melanie
Raizen, David M.
Fang-Yen, Christopher
Dehydrated Caenorhabditis elegans Stocks Are Resistant to Multiple Freeze-Thaw Cycles
title Dehydrated Caenorhabditis elegans Stocks Are Resistant to Multiple Freeze-Thaw Cycles
title_full Dehydrated Caenorhabditis elegans Stocks Are Resistant to Multiple Freeze-Thaw Cycles
title_fullStr Dehydrated Caenorhabditis elegans Stocks Are Resistant to Multiple Freeze-Thaw Cycles
title_full_unstemmed Dehydrated Caenorhabditis elegans Stocks Are Resistant to Multiple Freeze-Thaw Cycles
title_short Dehydrated Caenorhabditis elegans Stocks Are Resistant to Multiple Freeze-Thaw Cycles
title_sort dehydrated caenorhabditis elegans stocks are resistant to multiple freeze-thaw cycles
topic Investigations
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7718750/
https://www.ncbi.nlm.nih.gov/pubmed/33033066
http://dx.doi.org/10.1534/g3.120.401825
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