Isolated total RNA and protein are preserved after thawing for more than twenty-four hours

Isolated total RNA and protein are preserved after thawing for more than twenty-four hours

OBJECTIVE: The preservation of biological samples at a low temperature is important for later biochemical and/or histological analyses. However, the molecular viability of thawed samples has not been studied sufficiently in depth. The present study was undertaken to evaluate the viability of intact tissues, tissue homogenates, and isolated total RNA after defrosting for more than twenty-four hours. METHODS: The molecular viability of the thawed samples (n = 82) was assessed using the A260/A280 ratio, the RNA concentration, the RNA integrity, the level of intact mRNA determined by reverse transcriptase polymerase chain reaction, the protein level determined by Western blotting, and an examination of the histological structure. RESULTS: The integrity of the total RNA was not preserved in the thawed intact tissue, but the RNA integrity and level of mRNA were perfectly preserved in isolated defrosted samples of total RNA. Additionally, the level of β-actin protein was preserved in both thawed intact tissue and homogenates. CONCLUSION: Isolated total RNA does not undergo degradation due to thawing for at least 24 hours, and it is recommended to isolate the total RNA as soon as possible after tissue collection. Moreover, the protein level is preserved in defrosted tissues.

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Main Authors: Oliveira,Ivone Braga de, Ramos,Débora Rothstein, Lopes,Karen Lucasechi, Souza,Regiane Machado de, Heimann,Joel Claudio, Furukawa,Luzia Naôko Shinohara
Format: Digital revista
Language:English
Published: Faculdade de Medicina / USP 2012
Online Access:http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1807-59322012000300009
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spelling oai:scielo:S1807-593220120003000092012-03-27Isolated total RNA and protein are preserved after thawing for more than twenty-four hoursOliveira,Ivone Braga deRamos,Débora RothsteinLopes,Karen LucasechiSouza,Regiane Machado deHeimann,Joel ClaudioFurukawa,Luzia Naôko Shinohara Thawed tissue Defrosted isolated total RNA Ultra-low temperature Molecular viability Western blot OBJECTIVE: The preservation of biological samples at a low temperature is important for later biochemical and/or histological analyses. However, the molecular viability of thawed samples has not been studied sufficiently in depth. The present study was undertaken to evaluate the viability of intact tissues, tissue homogenates, and isolated total RNA after defrosting for more than twenty-four hours. METHODS: The molecular viability of the thawed samples (n = 82) was assessed using the A260/A280 ratio, the RNA concentration, the RNA integrity, the level of intact mRNA determined by reverse transcriptase polymerase chain reaction, the protein level determined by Western blotting, and an examination of the histological structure. RESULTS: The integrity of the total RNA was not preserved in the thawed intact tissue, but the RNA integrity and level of mRNA were perfectly preserved in isolated defrosted samples of total RNA. Additionally, the level of β-actin protein was preserved in both thawed intact tissue and homogenates. CONCLUSION: Isolated total RNA does not undergo degradation due to thawing for at least 24 hours, and it is recommended to isolate the total RNA as soon as possible after tissue collection. Moreover, the protein level is preserved in defrosted tissues.info:eu-repo/semantics/openAccessFaculdade de Medicina / USPClinics v.67 n.3 20122012-01-01info:eu-repo/semantics/articletext/htmlhttp://old.scielo.br/scielo.php?script=sci_arttext&pid=S1807-59322012000300009en10.6061/clinics/2012(03)09
institution SCIELO
collection OJS
country Brasil
countrycode BR
component Revista
access En linea
databasecode rev-scielo-br
tag revista
region America del Sur
libraryname SciELO
language English
format Digital
author Oliveira,Ivone Braga de
Ramos,Débora Rothstein
Lopes,Karen Lucasechi
Souza,Regiane Machado de
Heimann,Joel Claudio
Furukawa,Luzia Naôko Shinohara
spellingShingle Oliveira,Ivone Braga de
Ramos,Débora Rothstein
Lopes,Karen Lucasechi
Souza,Regiane Machado de
Heimann,Joel Claudio
Furukawa,Luzia Naôko Shinohara
Isolated total RNA and protein are preserved after thawing for more than twenty-four hours
author_facet Oliveira,Ivone Braga de
Ramos,Débora Rothstein
Lopes,Karen Lucasechi
Souza,Regiane Machado de
Heimann,Joel Claudio
Furukawa,Luzia Naôko Shinohara
author_sort Oliveira,Ivone Braga de
title Isolated total RNA and protein are preserved after thawing for more than twenty-four hours
title_short Isolated total RNA and protein are preserved after thawing for more than twenty-four hours
title_full Isolated total RNA and protein are preserved after thawing for more than twenty-four hours
title_fullStr Isolated total RNA and protein are preserved after thawing for more than twenty-four hours
title_full_unstemmed Isolated total RNA and protein are preserved after thawing for more than twenty-four hours
title_sort isolated total rna and protein are preserved after thawing for more than twenty-four hours
description OBJECTIVE: The preservation of biological samples at a low temperature is important for later biochemical and/or histological analyses. However, the molecular viability of thawed samples has not been studied sufficiently in depth. The present study was undertaken to evaluate the viability of intact tissues, tissue homogenates, and isolated total RNA after defrosting for more than twenty-four hours. METHODS: The molecular viability of the thawed samples (n = 82) was assessed using the A260/A280 ratio, the RNA concentration, the RNA integrity, the level of intact mRNA determined by reverse transcriptase polymerase chain reaction, the protein level determined by Western blotting, and an examination of the histological structure. RESULTS: The integrity of the total RNA was not preserved in the thawed intact tissue, but the RNA integrity and level of mRNA were perfectly preserved in isolated defrosted samples of total RNA. Additionally, the level of β-actin protein was preserved in both thawed intact tissue and homogenates. CONCLUSION: Isolated total RNA does not undergo degradation due to thawing for at least 24 hours, and it is recommended to isolate the total RNA as soon as possible after tissue collection. Moreover, the protein level is preserved in defrosted tissues.
publisher Faculdade de Medicina / USP
publishDate 2012
url http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1807-59322012000300009
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AT lopeskarenlucasechi isolatedtotalrnaandproteinarepreservedafterthawingformorethantwentyfourhours
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