Evidence map›Paper›PMID 38985280›Full record

ArticleMethods in molecular biology (Clifton, N.J.)2024

Quantification of DNA Methylation by ELISA in Epigenetic Studies in Plant Tissue Culture: A Theoretical-Practical Guide.

Marilia Santos Silva Patriota, Regina Beatriz Bernd, André Luis Xavier de Souza, Luís Alberto Martins Palhares de Melo, Jonny Everson Scherwinski-Pereira

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Article in Methods in molecular biology (Clifton, N.J.), 2024. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.

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4 · The record

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5 · Who and what money

Authors and funding

5 authors.

Marilia Santos Silva PatriotaEmbrapa Genetic Resources and Biotechnology, Brasília, DF, Brazil.
Regina Beatriz BerndEmbrapa Genetic Resources and Biotechnology, Brasília, DF, Brazil.
André Luis Xavier de SouzaEmbrapa Genetic Resources and Biotechnology, Brasília, DF, Brazil.
Luís Alberto Martins Palhares de MeloEmbrapa Genetic Resources and Biotechnology, Brasília, DF, Brazil.
Jonny Everson Scherwinski-PereiraEmbrapa Genetic Resources and Biotechnology, Brasília, DF, Brazil. jonny.pereira@embrapa.br.

Funding

No grant is acknowledged in the PubMed record.

6 · The paper itself

Abstract

This chapter describes a step-by-step protocol for rapid serological quantification of global DNA methylation by enzyme-linked immunosorbent assay (ELISA) in plant tissue culture specimens. As a case study model, we used the coconut palm (Cocos nucifera), from which plumules were subjected to somatic embryogenesis followed by embryogenic calli multiplication. DNA methylation is one of the most common epigenetic markers in the regulation of gene expression. DNA methylation is generally associated with non-expressed genes, that is, gene silencing under certain conditions, and the degree of DNA methylation can be used as a marker of various physiological processes, both in plants and in animal cells. Methylation consists of adding a methyl radical to carbon 5 of the DNA cytosine base. Herein, the global DNA methylation was quantified by ELISA with antibodies against methylated cytosines using a commercial kit (Zymo-Research™). The method allowed the detection of methylation in total DNA extracts from coconut palm embryogenic calli (arising from somatic embryogenesis) cultivated in liquid or solid media by using antibodies against methylated cytosines and enzymatic development with a colorimetric substrate. Control samples of commercially provided Escherichia coli bacterial DNA with previously known methylation percentages were included in the ELISA test to construct an experimental methylation standard curve. The logarithmic regression of this E. coli standard curve allowed methylation quantification in coconut palm samples. The present ELISA methodology, applied to coconut palm tissue culture specimens, is promising for use in other plant species and botanical families. This chapter is presented in a suitable format for use as a step-by-step laboratory procedure manual, with theoretical introduction information, which makes it easy to apply the protocol in samples of any biological nature to evaluate DNA global methylation associated with any physiological process.

Indexed as

DNA MethylationEnzyme-Linked Immunosorbent AssayEpigenesis, GeneticCocosDNA, PlantPlant Somatic Embryogenesis TechniquesTissue Culture TechniquesDNA, PlantDNA methylationELISAEpigeneticsPlant tissue culture

Identifiers

PMID38985280

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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.