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

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, ti...

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Bibliographic Details
Published in:Clinics (São Paulo, Brazil) Brazil), 2012-01, Vol.67 (3), p.255-259
Main Authors: de Oliveira, Ivone Braga, Ramos, Débora Rothstein, Lopes, Karen Lucasechi, de Souza, Regiane Machado, Heimann, Joel Claudio, Furukawa, Luzia Naôko Shinohara
Format: Article
Language:English
Subjects:
Actins - analysis
Animals
Basic Research
Cryopreservation - methods
Defrosted isolated total RNA
Gene Expression Profiling - methods
Male
MEDICINE, GENERAL & INTERNAL
Models, Animal
Molecular viability
Random Allocation
Rats
RNA - chemistry
RNA - genetics
RNA - isolation & purification
RNA Stability - genetics
RNA Stability - physiology
Specimen Handling - methods
Thawed tissue
Time Factors
Ultra-low temperature
Western blot
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Summary: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. 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. 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. 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.
ISSN:1807-5932
1980-5322
1980-5322
DOI:10.6061/clinics/2012(03)09