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Effect of preservation time of formalin-fixed paraffin-embedded tissues on extractable DNA and RNA quantity

OBJECTIVES: This study aimed to investigate the factors affecting the quantity of DNA and RNA extractable from human formalin-fixed paraffin-embedded (FFPE) tissues stored for different lengths of time. METHODS: We randomly selected 20 FFPE specimens harvested from hysteromyoma patients with uterine...

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Autores principales: Yi, Qing-qing, Yang, Rong, Shi, Jun-feng, Zeng, Nai-yan, Liang, Dong-yu, Sha, Shuang, Chang, Qing
Formato: Online Artículo Texto
Lenguaje:English
Publicado: SAGE Publications 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7309401/
https://www.ncbi.nlm.nih.gov/pubmed/32567435
http://dx.doi.org/10.1177/0300060520931259
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author Yi, Qing-qing
Yang, Rong
Shi, Jun-feng
Zeng, Nai-yan
Liang, Dong-yu
Sha, Shuang
Chang, Qing
author_facet Yi, Qing-qing
Yang, Rong
Shi, Jun-feng
Zeng, Nai-yan
Liang, Dong-yu
Sha, Shuang
Chang, Qing
author_sort Yi, Qing-qing
collection PubMed
description OBJECTIVES: This study aimed to investigate the factors affecting the quantity of DNA and RNA extractable from human formalin-fixed paraffin-embedded (FFPE) tissues stored for different lengths of time. METHODS: We randomly selected 20 FFPE specimens harvested from hysteromyoma patients with uterine fibroids during 2010, 2015, and 2017 at the Department of Pathology, Jiading District Central Hospital Affiliated Shanghai University of Medicine and Health Sciences. DNA and RNA extractions were performed using a DNA/RNA FFPE kit. DNA and RNA concentrations and their OD(260)/OD(280) ratios were determined by a NanoDrop 2000 spectrophotometer. The human β-globin gene and aldehyde dehydrogenase-2 (ALDH(2)) gene were amplified from nucleic acids using a LightCycler 480 Real-Time PCR System, and PCR amplification products were electrophoresed on 1% agarose gels. RESULTS: Specimens that were stored for longer showed more degradation and a reduced concentration of DNA and RNA after nucleic acid extraction. However, there was no significant difference in DNA or RNA purity. β-globin and ALDH(2) genes could be amplified from more than 99% of specimens. CONCLUSION: We found that FFPE tissues stored for longer had a reduced quantity of extractable DNA and RNA. However, these tissues could be used for the analysis of some small target genes.
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spelling pubmed-73094012020-06-30 Effect of preservation time of formalin-fixed paraffin-embedded tissues on extractable DNA and RNA quantity Yi, Qing-qing Yang, Rong Shi, Jun-feng Zeng, Nai-yan Liang, Dong-yu Sha, Shuang Chang, Qing J Int Med Res Retrospective Clinical Research Report OBJECTIVES: This study aimed to investigate the factors affecting the quantity of DNA and RNA extractable from human formalin-fixed paraffin-embedded (FFPE) tissues stored for different lengths of time. METHODS: We randomly selected 20 FFPE specimens harvested from hysteromyoma patients with uterine fibroids during 2010, 2015, and 2017 at the Department of Pathology, Jiading District Central Hospital Affiliated Shanghai University of Medicine and Health Sciences. DNA and RNA extractions were performed using a DNA/RNA FFPE kit. DNA and RNA concentrations and their OD(260)/OD(280) ratios were determined by a NanoDrop 2000 spectrophotometer. The human β-globin gene and aldehyde dehydrogenase-2 (ALDH(2)) gene were amplified from nucleic acids using a LightCycler 480 Real-Time PCR System, and PCR amplification products were electrophoresed on 1% agarose gels. RESULTS: Specimens that were stored for longer showed more degradation and a reduced concentration of DNA and RNA after nucleic acid extraction. However, there was no significant difference in DNA or RNA purity. β-globin and ALDH(2) genes could be amplified from more than 99% of specimens. CONCLUSION: We found that FFPE tissues stored for longer had a reduced quantity of extractable DNA and RNA. However, these tissues could be used for the analysis of some small target genes. SAGE Publications 2020-06-22 /pmc/articles/PMC7309401/ /pubmed/32567435 http://dx.doi.org/10.1177/0300060520931259 Text en © The Author(s) 2020 https://creativecommons.org/licenses/by-nc/4.0/ Creative Commons Non Commercial CC BY-NC: This article is distributed under the terms of the Creative Commons Attribution-NonCommercial 4.0 License (https://creativecommons.org/licenses/by-nc/4.0/) which permits non-commercial use, reproduction and distribution of the work without further permission provided the original work is attributed as specified on the SAGE and Open Access pages (https://us.sagepub.com/en-us/nam/open-access-at-sage).
spellingShingle Retrospective Clinical Research Report
Yi, Qing-qing
Yang, Rong
Shi, Jun-feng
Zeng, Nai-yan
Liang, Dong-yu
Sha, Shuang
Chang, Qing
Effect of preservation time of formalin-fixed paraffin-embedded tissues on extractable DNA and RNA quantity
title Effect of preservation time of formalin-fixed paraffin-embedded tissues on extractable DNA and RNA quantity
title_full Effect of preservation time of formalin-fixed paraffin-embedded tissues on extractable DNA and RNA quantity
title_fullStr Effect of preservation time of formalin-fixed paraffin-embedded tissues on extractable DNA and RNA quantity
title_full_unstemmed Effect of preservation time of formalin-fixed paraffin-embedded tissues on extractable DNA and RNA quantity
title_short Effect of preservation time of formalin-fixed paraffin-embedded tissues on extractable DNA and RNA quantity
title_sort effect of preservation time of formalin-fixed paraffin-embedded tissues on extractable dna and rna quantity
topic Retrospective Clinical Research Report
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7309401/
https://www.ncbi.nlm.nih.gov/pubmed/32567435
http://dx.doi.org/10.1177/0300060520931259
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