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Optimization of a Method for the Simultaneous Extraction of Polar and Non-Polar Oxylipin Metabolites, DNA, RNA, Small RNA, and Protein from a Single Small Tissue Sample

A more comprehensive picture of tissue biology can be obtained through the application and integration of multiple omic technologies. However, the common challenge in working with a precious sample is having a sample too small to separately extract analytes of interest for each experiment. Consideri...

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Autores principales: Hasegawa, Yu, Otoki, Yurika, McClorry, Shannon, Coates, Laurynne C., Lombardi, Rachel L., Taha, Ameer Y., Slupsky, Carolyn M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7564281/
https://www.ncbi.nlm.nih.gov/pubmed/32859006
http://dx.doi.org/10.3390/mps3030061
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author Hasegawa, Yu
Otoki, Yurika
McClorry, Shannon
Coates, Laurynne C.
Lombardi, Rachel L.
Taha, Ameer Y.
Slupsky, Carolyn M.
author_facet Hasegawa, Yu
Otoki, Yurika
McClorry, Shannon
Coates, Laurynne C.
Lombardi, Rachel L.
Taha, Ameer Y.
Slupsky, Carolyn M.
author_sort Hasegawa, Yu
collection PubMed
description A more comprehensive picture of tissue biology can be obtained through the application and integration of multiple omic technologies. However, the common challenge in working with a precious sample is having a sample too small to separately extract analytes of interest for each experiment. Considering the high heterogeneity that can be present in a single tissue sample, extracting all biomolecules from a single and undivided tissue is preferable because it allows direct comparison of results. Here, we combined a modified Folch extraction method with DNA, RNA, small RNA, and protein extraction using two commercial kits, which allowed us to extract polar metabolites and non-polar oxylipin metabolites, DNA, RNA, small RNA, and protein simultaneously from a small tissue sample. The method was validated in terms of quantity and quality of analytes for downstream analyses.
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spelling pubmed-75642812020-10-26 Optimization of a Method for the Simultaneous Extraction of Polar and Non-Polar Oxylipin Metabolites, DNA, RNA, Small RNA, and Protein from a Single Small Tissue Sample Hasegawa, Yu Otoki, Yurika McClorry, Shannon Coates, Laurynne C. Lombardi, Rachel L. Taha, Ameer Y. Slupsky, Carolyn M. Methods Protoc Protocol A more comprehensive picture of tissue biology can be obtained through the application and integration of multiple omic technologies. However, the common challenge in working with a precious sample is having a sample too small to separately extract analytes of interest for each experiment. Considering the high heterogeneity that can be present in a single tissue sample, extracting all biomolecules from a single and undivided tissue is preferable because it allows direct comparison of results. Here, we combined a modified Folch extraction method with DNA, RNA, small RNA, and protein extraction using two commercial kits, which allowed us to extract polar metabolites and non-polar oxylipin metabolites, DNA, RNA, small RNA, and protein simultaneously from a small tissue sample. The method was validated in terms of quantity and quality of analytes for downstream analyses. MDPI 2020-08-26 /pmc/articles/PMC7564281/ /pubmed/32859006 http://dx.doi.org/10.3390/mps3030061 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Protocol
Hasegawa, Yu
Otoki, Yurika
McClorry, Shannon
Coates, Laurynne C.
Lombardi, Rachel L.
Taha, Ameer Y.
Slupsky, Carolyn M.
Optimization of a Method for the Simultaneous Extraction of Polar and Non-Polar Oxylipin Metabolites, DNA, RNA, Small RNA, and Protein from a Single Small Tissue Sample
title Optimization of a Method for the Simultaneous Extraction of Polar and Non-Polar Oxylipin Metabolites, DNA, RNA, Small RNA, and Protein from a Single Small Tissue Sample
title_full Optimization of a Method for the Simultaneous Extraction of Polar and Non-Polar Oxylipin Metabolites, DNA, RNA, Small RNA, and Protein from a Single Small Tissue Sample
title_fullStr Optimization of a Method for the Simultaneous Extraction of Polar and Non-Polar Oxylipin Metabolites, DNA, RNA, Small RNA, and Protein from a Single Small Tissue Sample
title_full_unstemmed Optimization of a Method for the Simultaneous Extraction of Polar and Non-Polar Oxylipin Metabolites, DNA, RNA, Small RNA, and Protein from a Single Small Tissue Sample
title_short Optimization of a Method for the Simultaneous Extraction of Polar and Non-Polar Oxylipin Metabolites, DNA, RNA, Small RNA, and Protein from a Single Small Tissue Sample
title_sort optimization of a method for the simultaneous extraction of polar and non-polar oxylipin metabolites, dna, rna, small rna, and protein from a single small tissue sample
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7564281/
https://www.ncbi.nlm.nih.gov/pubmed/32859006
http://dx.doi.org/10.3390/mps3030061
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