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Test of the FlashFREEZE unit in tissue samples freezing for biobanking purposes

Availability of molecularly intact biospecimens is essential in genetic diagnostics to obtain credible results. Integrity of nucleic acids (particularly RNA) may be compromised at various steps of tissue handling, and affected by factors such as time to freeze, freezing technique and storing tempera...

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Autores principales: Biskup, Edyta, Schejbel, Lone, de Oliveira, Douglas Nogueira Perez, Høgdall, Estrid
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Springer Netherlands 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10209260/
https://www.ncbi.nlm.nih.gov/pubmed/36309911
http://dx.doi.org/10.1007/s10561-022-10045-1
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author Biskup, Edyta
Schejbel, Lone
de Oliveira, Douglas Nogueira Perez
Høgdall, Estrid
author_facet Biskup, Edyta
Schejbel, Lone
de Oliveira, Douglas Nogueira Perez
Høgdall, Estrid
author_sort Biskup, Edyta
collection PubMed
description Availability of molecularly intact biospecimens is essential in genetic diagnostics to obtain credible results. Integrity of nucleic acids (particularly RNA) may be compromised at various steps of tissue handling, and affected by factors such as time to freeze, freezing technique and storing temperature. At the same time, freezing and storing of the biological material should be feasible and safe for the operator. Here, we compared quality of DNA and RNA from biospecimens derived from different organs (breast, colon, adrenal glands, testes, rectum and uterus) frozen either using dry ice-cooled isopentane or with FlashFREEZE unit, in order to verify if the latter is suitable for routine use in biobanking. Implementing FlashFREEZE device would enable us to limit the use of isopentane, which is potentially toxic and environmentally harmful, whilst facilitate standardization of sample freezing time. We considered factors such RNA and DNA yield and purity. Furthermore, RNA integrity and RNA/DNA performance in routine analyses, such as qPCR, next generation sequencing or microarray, were also assessed. Our results indicate that freezing of tissue samples either with FlashFREEZE unit or isopentane ensures biological material with comparable expression profiles and DNA mutation status, indicating that RNA and DNA of similar quality can be extracted from both. Therefore, our findings support the use of the FlashFREEZE device in routine use for biobanking purposes.
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spelling pubmed-102092602023-05-26 Test of the FlashFREEZE unit in tissue samples freezing for biobanking purposes Biskup, Edyta Schejbel, Lone de Oliveira, Douglas Nogueira Perez Høgdall, Estrid Cell Tissue Bank Full Length Paper Availability of molecularly intact biospecimens is essential in genetic diagnostics to obtain credible results. Integrity of nucleic acids (particularly RNA) may be compromised at various steps of tissue handling, and affected by factors such as time to freeze, freezing technique and storing temperature. At the same time, freezing and storing of the biological material should be feasible and safe for the operator. Here, we compared quality of DNA and RNA from biospecimens derived from different organs (breast, colon, adrenal glands, testes, rectum and uterus) frozen either using dry ice-cooled isopentane or with FlashFREEZE unit, in order to verify if the latter is suitable for routine use in biobanking. Implementing FlashFREEZE device would enable us to limit the use of isopentane, which is potentially toxic and environmentally harmful, whilst facilitate standardization of sample freezing time. We considered factors such RNA and DNA yield and purity. Furthermore, RNA integrity and RNA/DNA performance in routine analyses, such as qPCR, next generation sequencing or microarray, were also assessed. Our results indicate that freezing of tissue samples either with FlashFREEZE unit or isopentane ensures biological material with comparable expression profiles and DNA mutation status, indicating that RNA and DNA of similar quality can be extracted from both. Therefore, our findings support the use of the FlashFREEZE device in routine use for biobanking purposes. Springer Netherlands 2022-10-30 2023 /pmc/articles/PMC10209260/ /pubmed/36309911 http://dx.doi.org/10.1007/s10561-022-10045-1 Text en © The Author(s) 2022 https://creativecommons.org/licenses/by/4.0/Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Full Length Paper
Biskup, Edyta
Schejbel, Lone
de Oliveira, Douglas Nogueira Perez
Høgdall, Estrid
Test of the FlashFREEZE unit in tissue samples freezing for biobanking purposes
title Test of the FlashFREEZE unit in tissue samples freezing for biobanking purposes
title_full Test of the FlashFREEZE unit in tissue samples freezing for biobanking purposes
title_fullStr Test of the FlashFREEZE unit in tissue samples freezing for biobanking purposes
title_full_unstemmed Test of the FlashFREEZE unit in tissue samples freezing for biobanking purposes
title_short Test of the FlashFREEZE unit in tissue samples freezing for biobanking purposes
title_sort test of the flashfreeze unit in tissue samples freezing for biobanking purposes
topic Full Length Paper
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10209260/
https://www.ncbi.nlm.nih.gov/pubmed/36309911
http://dx.doi.org/10.1007/s10561-022-10045-1
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